111224_PCR.xlsx

Selected Cell

Cell:

Value:

Raw XBP1 data Raw TRIB3 and HSPA5 data Processing Analyzed data Methods

Replicate and treatment are in the sample name column. 1 = replicate 1, 2 = replicate 2, 3 = replicate 3, m = replicate 4
NT = not treated, tun = tunicamycin treated, 4u = 4u8c treated
Replicates 1 and 2
Well	Well Position	Omit	Sample Name	Target Name	Task	Reporter	Quencher	RQ	RQ Min	RQ Max	CT	Ct Mean	Ct SD	Quantity	Delta Ct Mean	Delta Ct SD	Delta Delta Ct	Automatic Ct Threshold	Ct Threshold	Automatic Baseline	Baseline Start	Baseline End	Efficiency	Comments	Tm1	Tm2	Tm3	Custom1	Custom2	Custom3	Custom4	Custom5	Custom6	MTP
1	A1	false	1nt	xbp1 spliced	UNKNOWN	SYBR	None	1	0.8182626962661743	1.2221014499664307	30.288999557495117	30.415618896484375	0.10971656441688538		8.589860916137695	0.18051615357398987	0	true	0.10575682687542884	true	3	27	1		84.55587005615234									N
2	A2	false	1nt	xbp1 spliced	UNKNOWN	SYBR	None	1	0.8182626962661743	1.2221014499664307	30.476999282836914	30.415618896484375	0.10971656441688538		8.589860916137695	0.18051615357398987	0	true	0.10575682687542884	true	3	27	1		84.55587005615234									N
3	A3	false	1nt	xbp1 spliced	UNKNOWN	SYBR	None	1	0.8182626962661743	1.2221014499664307	30.481000900268555	30.415618896484375	0.10971656441688538		8.589860916137695	0.18051615357398987	0	true	0.10575682687542884	true	3	27	1		84.55587005615234									N
7	A7	false	2nt	xbp1 spliced	UNKNOWN	SYBR	None	0.9528027772903442	0.8740479946136475	1.0386537313461304	30.617000579833984	30.56184959411621	0.06257332861423492		8.659611701965332	0.07764605432748795	0.06975046545267105	true	0.10575682687542884	true	3	27	1		84.75274658203125									N
8	A8	false	2nt	xbp1 spliced	UNKNOWN	SYBR	None	0.9528027772903442	0.8740479946136475	1.0386537313461304	30.493999481201172	30.56184959411621	0.06257332861423492		8.659611701965332	0.07764605432748795	0.06975046545267105	true	0.10575682687542884	true	3	27	1		84.65430450439453									N
9	A9	false	2nt	xbp1 spliced	UNKNOWN	SYBR	None	0.9528027772903442	0.8740479946136475	1.0386537313461304	30.575000762939453	30.56184959411621	0.06257332861423492		8.659611701965332	0.07764605432748795	0.06975046545267105	true	0.10575682687542884	true	3	27	1		84.55587005615234									N
49	E1	false	1tun	xbp1 spliced	UNKNOWN	SYBR	None	145.83819580078125	141.48167419433594	150.328857421875	24.365999221801758	24.386056900024414	0.017452871426939964		1.4016361236572266	0.02729501947760582	-7.188224792480469	true	0.10575682687542884	true	3	20	1		84.55587005615234									N
50	E2	false	1tun	xbp1 spliced	UNKNOWN	SYBR	None	145.83819580078125	141.48167419433594	150.328857421875	24.39299964904785	24.386056900024414	0.017452871426939964		1.4016361236572266	0.02729501947760582	-7.188224792480469	true	0.10575682687542884	true	3	20	1		84.65430450439453									N
51	E3	false	1tun	xbp1 spliced	UNKNOWN	SYBR	None	145.83819580078125	141.48167419433594	150.328857421875	24.39900016784668	24.386056900024414	0.017452871426939964		1.4016361236572266	0.02729501947760582	-7.188224792480469	true	0.10575682687542884	true	3	20	1		84.55587005615234									N
55	E7	false	2tun	xbp1 spliced	UNKNOWN	SYBR	None	204.15867614746094	176.13702392578125	236.6383056640625	22.954999923706055	22.971158981323242	0.037734150886535645		0.9163138270378113	0.13287286460399628	-7.673547267913818	true	0.10575682687542884	true	3	19	1		84.55587005615234									N
56	E8	false	2tun	xbp1 spliced	UNKNOWN	SYBR	None	204.15867614746094	176.13702392578125	236.6383056640625	22.944000244140625	22.971158981323242	0.037734150886535645		0.9163138270378113	0.13287286460399628	-7.673547267913818	true	0.10575682687542884	true	3	18	1		84.45743560791016									N
57	E9	false	2tun	xbp1 spliced	UNKNOWN	SYBR	None	204.15867614746094	176.13702392578125	236.6383056640625	23.013999938964844	22.971158981323242	0.037734150886535645		0.9163138270378113	0.13287286460399628	-7.673547267913818	true	0.10575682687542884	true	3	19	1		84.45743560791016									N
85	H1	false	1tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.052415881305933	0.03492182493209839	0.07867357134819031	32.9630012512207	33.34567642211914	0.36268433928489685		12.84371280670166	0.3654909133911133	4.253852367401123	true	0.10575682687542884	true	3	28	1		84.35900115966797									N
86	H2	false	1tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.052415881305933	0.03492182493209839	0.07867357134819031	33.388999938964844	33.34567642211914	0.36268433928489685		12.84371280670166	0.3654909133911133	4.253852367401123	true	0.10575682687542884	true	3	28	1		84.45743560791016									N
87	H3	false	1tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.052415881305933	0.03492182493209839	0.07867357134819031	33.685001373291016	33.34567642211914	0.36268433928489685		12.84371280670166	0.3654909133911133	4.253852367401123	true	0.10575682687542884	true	3	28	1		84.45743560791016									N
91	H7	false	2tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.035957515239715576	0.021047404035925865	0.06143003702163696	35.50699996948242	35.02152633666992	0.4690195322036743		13.387423515319824	0.4820087254047394	4.797563076019287	true	0.10575682687542884	true	3	29	1		84.65430450439453									N
92	H8	false	2tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.035957515239715576	0.021047404035925865	0.06143003702163696	34.98699951171875	35.02152633666992	0.4690195322036743		13.387423515319824	0.4820087254047394	4.797563076019287	true	0.10575682687542884	true	3	29	1		84.55587005615234									N
93	H9	false	2tun4u	xbp1 spliced	UNKNOWN	SYBR	None	0.035957515239715576	0.021047404035925865	0.06143003702163696	34.57099914550781	35.02152633666992	0.4690195322036743		13.387423515319824	0.4820087254047394	4.797563076019287	true	0.10575682687542884	true	3	29	1		84.55587005615234									N
4	A4	false	1nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.929000854492188	21.825759887695312	0.14334698021411896					true	0.1751817209411774	true	3	17	1		83.96526336669922									N
5	A5	false	1nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.88599967956543	21.825759887695312	0.14334698021411896					true	0.1751817209411774	true	3	17	1		83.96526336669922									N
6	A6	false	1nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.66200065612793	21.825759887695312	0.14334698021411896					true	0.1751817209411774	true	3	16	1		84.0636978149414									N
10	A10	false	2nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.937999725341797	21.902236938476562	0.04597269371151924					true	0.1751817209411774	true	3	16	1		83.86682891845703									N
11	A11	false	2nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.91900062561035	21.902236938476562	0.04597269371151924					true	0.1751817209411774	true	3	16	1		83.86682891845703									N
12	A12	false	2nt	xbp1 unspliced	UNKNOWN	SYBR	None				21.850000381469727	21.902236938476562	0.04597269371151924					true	0.1751817209411774	true	3	16	1		83.86682891845703									N
52	E4	false	1tun	xbp1 unspliced	UNKNOWN	SYBR	None				22.9950008392334	22.984420776367188	0.020986076444387436					true	0.1751817209411774	true	3	18	1		83.96526336669922									N
53	E5	false	1tun	xbp1 unspliced	UNKNOWN	SYBR	None				22.99799919128418	22.984420776367188	0.020986076444387436					true	0.1751817209411774	true	3	18	1		83.96526336669922									N
54	E6	false	1tun	xbp1 unspliced	UNKNOWN	SYBR	None				22.959999084472656	22.984420776367188	0.020986076444387436					true	0.1751817209411774	true	3	18	1		83.96526336669922									N
58	E10	false	2tun	xbp1 unspliced	UNKNOWN	SYBR	None				22.104000091552734	22.054845809936523	0.12740224599838257					true	0.1751817209411774	true	3	17	1		83.86682891845703									N
59	E11	false	2tun	xbp1 unspliced	UNKNOWN	SYBR	None				22.149999618530273	22.054845809936523	0.12740224599838257					true	0.1751817209411774	true	3	17	1		83.86682891845703									N
60	E12	false	2tun	xbp1 unspliced	UNKNOWN	SYBR	None				21.90999984741211	22.054845809936523	0.12740224599838257					true	0.1751817209411774	true	3	16	1		83.96526336669922									N
88	H4	false	1tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				20.545000076293945	20.50196647644043	0.045207127928733826					true	0.1751817209411774	true	3	15	1		83.86682891845703									N
89	H5	false	1tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				20.5049991607666	20.50196647644043	0.045207127928733826					true	0.1751817209411774	true	3	15	1		83.86682891845703									N
90	H6	false	1tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				20.454999923706055	20.50196647644043	0.045207127928733826					true	0.1751817209411774	true	3	15	1		83.96526336669922									N
94	H10	false	2tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				21.594999313354492	21.634103775024414	0.11114447563886642					true	0.1751817209411774	true	3	16	1		83.76839447021484									N
95	H11	false	2tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				21.54800033569336	21.634103775024414	0.11114447563886642					true	0.1751817209411774	true	3	16	1		83.66996002197266									N
96	H12	false	2tun4u	xbp1 unspliced	UNKNOWN	SYBR	None				21.759000778198242	21.634103775024414	0.11114447563886642					true	0.1751817209411774	true	3	17	1		83.66996002197266									N

Replicates 3 and 4
Well	Well Position	Omit	Sample Name	Target Name	Task	Reporter	Quencher	RQ	RQ Min	RQ Max	CT	Ct Mean	Ct SD	Quantity	Delta Ct Mean	Delta Ct SD	Delta Delta Ct	Automatic Ct Threshold	Ct Threshold	Automatic Baseline	Baseline Start	Baseline End	Efficiency	Comments	Tm1	Tm2	Tm3	Custom1	Custom2	Custom3	Custom4	Custom5	Custom6	HIGHSD	MTP
1	A1	false	3nt	xbp1 sp	UNKNOWN	SYBR	None	1	0.930397629737854	1.0748093128204346	32.29600143432617	32.285579681396484	0.061162084341049194		8.322842597961426	0.06492944806814194	0	true	0.11800127860501854	true	3	27	1		84.56365966796875									N	N
2	A2	false	3nt	xbp1 sp	UNKNOWN	SYBR	None	1	0.930397629737854	1.0748093128204346	32.340999603271484	32.285579681396484	0.061162084341049194		8.322842597961426	0.06492944806814194	0	true	0.11800127860501854	true	3	27	1		84.56365966796875									N	N
3	A3	false	3nt	xbp1 sp	UNKNOWN	SYBR	None	1	0.930397629737854	1.0748093128204346	32.220001220703125	32.285579681396484	0.061162084341049194		8.322842597961426	0.06492944806814194	0	true	0.11800127860501854	true	3	27	1		84.56365966796875									N	N
7	A7	false	mnt	xbp1 sp	UNKNOWN	SYBR	None	1.2887864112854004	1.072513461112976	1.548671007156372	30.6560001373291	30.83660888671875	0.15729603171348572		7.95682954788208	0.16532787680625916	-0.36601319909095764	true	0.11800127860501854	true	3	26	1		84.66212463378906									N	N
8	A8	false	mnt	xbp1 sp	UNKNOWN	SYBR	None	1.2887864112854004	1.072513461112976	1.548671007156372	30.94099998474121	30.83660888671875	0.15729603171348572		7.95682954788208	0.16532787680625916	-0.36601319909095764	true	0.11800127860501854	true	3	25	1		84.56365966796875									N	N
9	A9	false	mnt	xbp1 sp	UNKNOWN	SYBR	None	1.2887864112854004	1.072513461112976	1.548671007156372	30.913000106811523	30.83660888671875	0.15729603171348572		7.95682954788208	0.16532787680625916	-0.36601319909095764	true	0.11800127860501854	true	3	26	1		84.56365966796875									N	N
49	E1	false	3tun	xbp1 sp	UNKNOWN	SYBR	None	236.30685424804688	219.14183044433594	254.81637573242188	23.638999938964844	23.601655960083008	0.040742628276348114		0.43832525610923767	0.06787138432264328	-7.884517669677734	true	0.11800127860501854	true	3	19	1		84.66212463378906									N	N
50	E2	false	3tun	xbp1 sp	UNKNOWN	SYBR	None	236.30685424804688	219.14183044433594	254.81637573242188	23.558000564575195	23.601655960083008	0.040742628276348114		0.43832525610923767	0.06787138432264328	-7.884517669677734	true	0.11800127860501854	true	3	19	1		84.76058959960938									N	N
51	E3	false	3tun	xbp1 sp	UNKNOWN	SYBR	None	236.30685424804688	219.14183044433594	254.81637573242188	23.60700035095215	23.601655960083008	0.040742628276348114		0.43832525610923767	0.06787138432264328	-7.884517669677734	true	0.11800127860501854	true	3	19	1		84.66212463378906									N	N
55	E7	false	mtun	xbp1 sp	UNKNOWN	SYBR	None	231.7190704345703	214.57675170898438	250.23086547851562	23.3799991607666	23.342636108398438	0.04997463524341583		0.4666099548339844	0.06917297840118408	-7.8562331199646	true	0.11800127860501854	true	3	18	1		84.56365966796875									N	N
56	E8	false	mtun	xbp1 sp	UNKNOWN	SYBR	None	231.7190704345703	214.57675170898438	250.23086547851562	23.363000869750977	23.342636108398438	0.04997463524341583		0.4666099548339844	0.06917297840118408	-7.8562331199646	true	0.11800127860501854	true	3	19	1		84.56365966796875									N	N
57	E9	false	mtun	xbp1 sp	UNKNOWN	SYBR	None	231.7190704345703	214.57675170898438	250.23086547851562	23.285999298095703	23.342636108398438	0.04997463524341583		0.4666099548339844	0.06917297840118408	-7.8562331199646	true	0.11800127860501854	true	3	18	1		84.46520233154297									N	N
85	H1	false	3tun4uc	xbp1 sp	UNKNOWN	SYBR	None	0.03784313052892685	0.029094120487570763	0.04922308772802353	34.979000091552734	34.864803314208984	0.23662219941616058		13.04666805267334	0.23662395775318146	4.723824977874756	true	0.11800127860501854	true	3	30	1		84.36673736572266									N	N
86	H2	false	3tun4uc	xbp1 sp	UNKNOWN	SYBR	None	0.03784313052892685	0.029094120487570763	0.04922308772802353	34.59299850463867	34.864803314208984	0.23662219941616058		13.04666805267334	0.23662395775318146	4.723824977874756	true	0.11800127860501854	true	3	29	1		84.36673736572266									N	N
87	H3	false	3tun4uc	xbp1 sp	UNKNOWN	SYBR	None	0.03784313052892685	0.029094120487570763	0.04922308772802353	35.02299880981445	34.864803314208984	0.23662219941616058		13.04666805267334	0.23662395775318146	4.723824977874756	true	0.11800127860501854	true	3	30	1		84.46520233154297									N	N
91	H7	false	mtun4u	xbp1 sp	UNKNOWN	SYBR	None	0.040832120925188065	0.0209805928170681	0.07946687191724777	36.4739990234375	37.14285659790039	0.5987184643745422		12.936994552612305	0.5992890000343323	4.614151477813721	true	0.11800127860501854	true	3	31	1		84.66212463378906									Y	N
92	H8	false	mtun4u	xbp1 sp	UNKNOWN	SYBR	None	0.040832120925188065	0.0209805928170681	0.07946687191724777	37.32500076293945	37.14285659790039	0.5987184643745422		12.936994552612305	0.5992890000343323	4.614151477813721	true	0.11800127860501854	true	3	32	1		84.56365966796875									Y	N
93	H9	false	mtun4u	xbp1 sp	UNKNOWN	SYBR	None	0.040832120925188065	0.0209805928170681	0.07946687191724777	37.62900161743164	37.14285659790039	0.5987184643745422		12.936994552612305	0.5992890000343323	4.614151477813721	true	0.11800127860501854	true	3	32	1		84.56365966796875									Y	N
4	A4	false	3nt	xbp1 uns	UNKNOWN	SYBR	None				23.986000061035156	23.962738037109375	0.021795254200696945					true	0.257616	true	3	18	1		83.97288513183594									N	N
5	A5	false	3nt	xbp1 uns	UNKNOWN	SYBR	None				23.959999084472656	23.962738037109375	0.021795254200696945					true	0.257616	true	3	18	1		83.97288513183594									N	N
6	A6	false	3nt	xbp1 uns	UNKNOWN	SYBR	None				23.941999435424805	23.962738037109375	0.021795254200696945					true	0.257616	true	3	18	1		84.07135009765625									N	N
10	A10	false	mnt	xbp1 uns	UNKNOWN	SYBR	None				22.8799991607666	22.879777908325195	0.05090446025133133					true	0.257616	true	3	17	1		83.87442016601562									N	N
11	A11	false	mnt	xbp1 uns	UNKNOWN	SYBR	None				22.930999755859375	22.879777908325195	0.05090446025133133					true	0.257616	true	3	17	1		83.87442016601562									N	N
12	A12	false	mnt	xbp1 uns	UNKNOWN	SYBR	None				22.82900047302246	22.879777908325195	0.05090446025133133					true	0.257616	true	3	17	1		83.77595520019531									N	N
52	E4	false	3tun	xbp1 uns	UNKNOWN	SYBR	None				23.204999923706055	23.163331985473633	0.054282255470752716					true	0.257616	true	3	17	1		83.97288513183594									N	N
53	E5	false	3tun	xbp1 uns	UNKNOWN	SYBR	None				23.18199920654297	23.163331985473633	0.054282255470752716					true	0.257616	true	3	17	1		83.97288513183594									N	N
54	E6	false	3tun	xbp1 uns	UNKNOWN	SYBR	None				23.101999282836914	23.163331985473633	0.054282255470752716					true	0.257616	true	3	17	1		83.97288513183594									N	N
58	E10	false	mtun	xbp1 uns	UNKNOWN	SYBR	None				22.899999618530273	22.876028060913086	0.04782715067267418					true	0.257616	true	3	17	1		83.87442016601562									N	N
59	E11	false	mtun	xbp1 uns	UNKNOWN	SYBR	None				22.906999588012695	22.876028060913086	0.04782715067267418					true	0.257616	true	3	17	1		83.87442016601562									N	N
60	E12	false	mtun	xbp1 uns	UNKNOWN	SYBR	None				22.820999145507812	22.876028060913086	0.04782715067267418					true	0.257616	true	3	17	1		83.87442016601562									N	N
88	H4	false	3tun4uc	xbp1 uns	UNKNOWN	SYBR	None				21.81800079345703	21.81813621520996	9.115736465901136E-4					true	0.257616	true	3	16	1		83.77595520019531									N	N
89	H5	false	3tun4uc	xbp1 uns	UNKNOWN	SYBR	None				21.816999435424805	21.81813621520996	9.115736465901136E-4					true	0.257616	true	3	16	1		83.87442016601562									N	N
90	H6	false	3tun4uc	xbp1 uns	UNKNOWN	SYBR	None				21.819000244140625	21.81813621520996	9.115736465901136E-4					true	0.257616	true	3	17	1		83.87442016601562									N	N
94	H10	false	mtun4u	xbp1 uns	UNKNOWN	SYBR	None				24.18600082397461	24.20586395263672	0.026144949719309807					true	0.257616	true	3	18	1		83.67749786376953									N	N
95	H11	false	mtun4u	xbp1 uns	UNKNOWN	SYBR	None				24.195999145507812	24.20586395263672	0.026144949719309807					true	0.257616	true	3	18	1		83.57903289794922									N	N
96	H12	false	mtun4u	xbp1 uns	UNKNOWN	SYBR	None				24.236000061035156	24.20586395263672	0.026144949719309807					true	0.257616	true	3	18	1		83.57903289794922									N	N

100

Replicates are labelled at the top
This info exported directly from the plate reader.




Replicate 1
Sample Name	Target Name	Task	Reporter	Quencher	RQ	RQ Min	RQ Max	CT	Ct Mean	Ct SD	Quantity	Delta Ct Mean	Delta Ct SD	Delta Delta Ct	Automatic Ct Threshold	Ct Threshold	Automatic Baseline	Baseline Start	Baseline End	Efficiency	Comments	Tm1	Tm2	Tm3	Custom1	Custom2	Custom3	Custom4	Custom5	Custom6
rnt	actin	UNKNOWN	SYBR	None				23.878000259399414	23.81490135192871	0.05811982601881027					true	0.28577011490197257	true	3	17	1		90.40977478027344
rnt	actin	UNKNOWN	SYBR	None				23.804000854492188	23.81490135192871	0.05811982601881027					true	0.28577011490197257	true	3	17	1		90.40977478027344
rnt	actin	UNKNOWN	SYBR	None				23.76300048828125	23.81490135192871	0.05811982601881027					true	0.28577011490197257	true	3	17	1		90.60696411132812
rnt	hspa5	UNKNOWN	SYBR	None	1	0.9303392767906189	1.0748766660690308	20.871999740600586	20.90586280822754	0.029073234647512436		-2.9090373516082764	0.06498590111732483	0	true	0.255036	true	3	15	1		86.564697265625
rnt	hspa5	UNKNOWN	SYBR	None	1	0.9303392767906189	1.0748766660690308	20.92099952697754	20.90586280822754	0.029073234647512436		-2.9090373516082764	0.06498590111732483	0	true	0.255036	true	3	15	1		86.46611022949219
rnt	hspa5	UNKNOWN	SYBR	None	1	0.9303392767906189	1.0748766660690308	20.923999786376953	20.90586280822754	0.029073234647512436		-2.9090373516082764	0.06498590111732483	0	true	0.255036	true	3	15	1		86.46611022949219
rnt	trib3	UNKNOWN	SYBR	None	1	0.851277232170105	1.1747053861618042	28.236000061035156	28.112733840942383	0.13275159895420074		4.297834873199463	0.14491687715053558	0	true	0.23067519326743996	true	3	23	1		84.79004669189453
rnt	trib3	UNKNOWN	SYBR	None	1	0.851277232170105	1.1747053861618042	28.1299991607666	28.112733840942383	0.13275159895420074		4.297834873199463	0.14491687715053558	0	true	0.23067519326743996	true	3	22	1		84.69145202636719
rnt	trib3	UNKNOWN	SYBR	None	1	0.851277232170105	1.1747053861618042	27.972000122070312	28.112733840942383	0.13275159895420074		4.297834873199463	0.14491687715053558	0	true	0.23067519326743996	true	3	22	1		84.69145202636719
rtun	actin	UNKNOWN	SYBR	None				23.61400032043457	23.559701919555664	0.05114906281232834					true	0.28577011490197257	true	3	17	1		90.40977478027344
rtun	actin	UNKNOWN	SYBR	None				23.552000045776367	23.559701919555664	0.05114906281232834					true	0.28577011490197257	true	3	17	1		90.40977478027344
rtun	actin	UNKNOWN	SYBR	None				23.51300048828125	23.559701919555664	0.05114906281232834					true	0.28577011490197257	true	3	17	1		90.40977478027344
rtun	hspa5	UNKNOWN	SYBR	None	19.777463912963867	18.676197052001953	20.943666458129883	16.34600067138672	16.344879150390625	0.0065308380872011185		-7.214822769165039	0.05156431347131729	-4.305785655975342	true	0.255036	true	3	11	1		86.46611022949219
rtun	hspa5	UNKNOWN	SYBR	None	19.777463912963867	18.676197052001953	20.943666458129883	16.33799934387207	16.344879150390625	0.0065308380872011185		-7.214822769165039	0.05156431347131729	-4.305785655975342	true	0.255036	true	3	11	1		86.46611022949219
rtun	hspa5	UNKNOWN	SYBR	None	19.777463912963867	18.676197052001953	20.943666458129883	16.35099983215332	16.344879150390625	0.0065308380872011185		-7.214822769165039	0.05156431347131729	-4.305785655975342	true	0.255036	true	3	11	1		86.46611022949219
rtun	trib3	UNKNOWN	SYBR	None	46.85881042480469	43.9566650390625	49.95256805419922	22.299999237060547	22.307289123535156	0.026359744369983673		-1.2524133920669556	0.05754183232784271	-5.550248622894287	true	0.23067519326743996	true	3	17	1		84.69145202636719
rtun	trib3	UNKNOWN	SYBR	None	46.85881042480469	43.9566650390625	49.95256805419922	22.28499984741211	22.307289123535156	0.026359744369983673		-1.2524133920669556	0.05754183232784271	-5.550248622894287	true	0.23067519326743996	true	3	17	1		84.69145202636719
rtun	trib3	UNKNOWN	SYBR	None	46.85881042480469	43.9566650390625	49.95256805419922	22.336999893188477	22.307289123535156	0.026359744369983673		-1.2524133920669556	0.05754183232784271	-5.550248622894287	true	0.23067519326743996	true	3	17	1		84.59286499023438
rtunca7	actin	UNKNOWN	SYBR	None				27.242000579833984	27.151601791381836	0.0790543332695961					true	0.28577011490197257	true	3	21	1		90.50837707519531
rtunca7	actin	UNKNOWN	SYBR	None				27.09600067138672	27.151601791381836	0.0790543332695961					true	0.28577011490197257	true	3	21	1		90.40977478027344
rtunca7	actin	UNKNOWN	SYBR	None				27.117000579833984	27.151601791381836	0.0790543332695961					true	0.28577011490197257	true	3	20	1		90.40977478027344
rtunca7	hspa5	UNKNOWN	SYBR	None	4.831404209136963	4.422825336456299	5.277727127075195	21.972999572753906	21.970123291015625	0.008622989989817142		-5.1814799308776855	0.0795232281088829	-2.272442579269409	true	0.255036	true	3	16	1		86.564697265625
rtunca7	hspa5	UNKNOWN	SYBR	None	4.831404209136963	4.422825336456299	5.277727127075195	21.959999084472656	21.970123291015625	0.008622989989817142		-5.1814799308776855	0.0795232281088829	-2.272442579269409	true	0.255036	true	3	16	1		86.46611022949219
rtunca7	hspa5	UNKNOWN	SYBR	None	4.831404209136963	4.422825336456299	5.277727127075195	21.976999282836914	21.970123291015625	0.008622989989817142		-5.1814799308776855	0.0795232281088829	-2.272442579269409	true	0.255036	true	3	16	1		86.46611022949219
rtungsk	actin	UNKNOWN	SYBR	None				24.31399917602539	24.247053146362305	0.06766768544912338					true	0.28577011490197257	true	3	18	1		90.31118774414062
rtungsk	actin	UNKNOWN	SYBR	None				24.24799919128418	24.247053146362305	0.06766768544912338					true	0.28577011490197257	true	3	18	1		90.31118774414062
rtungsk	actin	UNKNOWN	SYBR	None				24.179000854492188	24.247053146362305	0.06766768544912338					true	0.28577011490197257	true	3	18	1		90.31118774414062
rtungsk	trib3	UNKNOWN	SYBR	None	3.192003011703491	2.7249226570129395	3.7391457557678223	26.90999984741211	26.87042808532715	0.1252823770046234		2.623373031616211	0.14238886535167694	-1.6744619607925415	true	0.23067519326743996	true	3	21	1		84.69145202636719
rtungsk	trib3	UNKNOWN	SYBR	None	3.192003011703491	2.7249226570129395	3.7391457557678223	26.97100067138672	26.87042808532715	0.1252823770046234		2.623373031616211	0.14238886535167694	-1.6744619607925415	true	0.23067519326743996	true	3	21	1		84.59286499023438
rtungsk	trib3	UNKNOWN	SYBR	None	3.192003011703491	2.7249226570129395	3.7391457557678223	26.729999542236328	26.87042808532715	0.1252823770046234		2.623373031616211	0.14238886535167694	-1.6744619607925415	true	0.23067519326743996	true	3	21	1		84.59286499023438

Replicate 2					0.06811959123089062
1nt	actin	UNKNOWN	SYBR	None				27.552000045776367	27.796545028686523	0.25439295172691345					true	0.19068063569905777	true	3	22	1		90.25914001464844									N	N	N	N
1nt	actin	UNKNOWN	SYBR	None				27.77899932861328	27.796545028686523	0.25439295172691345					true	0.19068063569905777	true	3	22	1		90.25914001464844									N	N	N	N
1nt	actin	UNKNOWN	SYBR	None				28.05900001525879	27.796545028686523	0.25439295172691345					true	0.19068063569905777	true	3	22	1		90.35723876953125									N	N	N	N
1nt	hspa5	UNKNOWN	SYBR	None	1	0.7532291412353516	1.3276172876358032	22.8439998626709	22.859365463256836	0.018286043778061867		-4.9371795654296875	0.25504931807518005	0	true	0.2005121203624967	true	3	17	1		86.43318176269531									N	N	N	N
1nt	hspa5	UNKNOWN	SYBR	None	1	0.7532291412353516	1.3276172876358032	22.8799991607666	22.859365463256836	0.018286043778061867		-4.9371795654296875	0.25504931807518005	0	true	0.2005121203624967	true	3	17	1		86.43318176269531									N	N	N	N
1nt	hspa5	UNKNOWN	SYBR	None	1	0.7532291412353516	1.3276172876358032	22.854000091552734	22.859365463256836	0.018286043778061867		-4.9371795654296875	0.25504931807518005	0	true	0.2005121203624967	true	3	17	1		86.53128051757812									N	N	N	N
1nt	trib3	UNKNOWN	SYBR	None	1	0.2697068154811859	3.7077300548553467	34.12699890136719	32.847896575927734	1.1516246795654297		5.051351070404053	1.1793875694274902	0	true	0.19439522035735243	true	3	29	1		84.47115325927734	79.07557678222656								Y	N	N	Y
1nt	trib3	UNKNOWN	SYBR	None	1	0.2697068154811859	3.7077300548553467	32.52199935913086	32.847896575927734	1.1516246795654297		5.051351070404053	1.1793875694274902	0	true	0.19439522035735243	true	3	26	1		84.56925201416016									Y	N	N	N
1nt	trib3	UNKNOWN	SYBR	None	1	0.2697068154811859	3.7077300548553467	31.893999099731445	32.847896575927734	1.1516246795654297		5.051351070404053	1.1793875694274902	0	true	0.19439522035735243	true	3	26	1		84.56925201416016									Y	N	N	N
1tun	hspa5	UNKNOWN	SYBR	None	5.9979047775268555	5.1767802238464355	6.949274063110352	18.659000396728516	18.612991333007812	0.05307963117957115		-7.5216383934021	0.1325053572654724	-2.584458589553833	true	0.2005121203624967	true	3	14	1		86.43318176269531									N	N	N	N
1tun	hspa5	UNKNOWN	SYBR	None	5.9979047775268555	5.1767802238464355	6.949274063110352	18.55500030517578	18.612991333007812	0.05307963117957115		-7.5216383934021	0.1325053572654724	-2.584458589553833	true	0.2005121203624967	true	3	13	1		86.43318176269531									N	N	N	N
1tun	hspa5	UNKNOWN	SYBR	None	5.9979047775268555	5.1767802238464355	6.949274063110352	18.625999450683594	18.612991333007812	0.05307963117957115		-7.5216383934021	0.1325053572654724	-2.584458589553833	true	0.2005121203624967	true	3	13	1		86.43318176269531									N	N	N	N
1tun	actin	UNKNOWN	SYBR	None				26.049999237060547	26.13463020324707	0.1214093267917633					true	0.19068063569905777	true	3	20	1		90.45533752441406									N	N	N	N
1tun	actin	UNKNOWN	SYBR	None				26.08099937438965	26.13463020324707	0.1214093267917633					true	0.19068063569905777	true	3	20	1		90.45533752441406									N	N	N	N
1tun	actin	UNKNOWN	SYBR	None				26.27400016784668	26.13463020324707	0.1214093267917633					true	0.19068063569905777	true	3	20	1		90.45533752441406									N	N	N	N
1tun	trib3	UNKNOWN	SYBR	None	81.77969360351562	71.09056091308594	94.07603454589844	24.816999435424805	24.832311630249023	0.03395451232790947		-1.302319884300232	0.12606796622276306	-6.353670597076416	true	0.19439522035735243	true	3	19	1		84.56925201416016									N	N	N	N
1tun	trib3	UNKNOWN	SYBR	None	81.77969360351562	71.09056091308594	94.07603454589844	24.80900001525879	24.832311630249023	0.03395451232790947		-1.302319884300232	0.12606796622276306	-6.353670597076416	true	0.19439522035735243	true	3	19	1		84.56925201416016									N	N	N	N
1tun	trib3	UNKNOWN	SYBR	None	81.77969360351562	71.09056091308594	94.07603454589844	24.871000289916992	24.832311630249023	0.03395451232790947		-1.302319884300232	0.12606796622276306	-6.353670597076416	true	0.19439522035735243	true	3	19	1		84.56925201416016									N	N	N	N
1tunca7	hspa5	UNKNOWN	SYBR	None	2.0208442211151123	1.4474107027053833	2.8214597702026367	21.5	21.481016159057617	0.01815144531428814		-5.952137470245361	0.30036765336990356	-1.0149580240249634	true	0.2005121203624967	true	3	16	1		86.43318176269531									N	N	N	N
1tunca7	hspa5	UNKNOWN	SYBR	None	2.0208442211151123	1.4474107027053833	2.8214597702026367	21.464000701904297	21.481016159057617	0.01815144531428814		-5.952137470245361	0.30036765336990356	-1.0149580240249634	true	0.2005121203624967	true	3	16	1		86.43318176269531									N	N	N	N
1tunca7	hspa5	UNKNOWN	SYBR	None	2.0208442211151123	1.4474107027053833	2.8214597702026367	21.479000091552734	21.481016159057617	0.01815144531428814		-5.952137470245361	0.30036765336990356	-1.0149580240249634	true	0.2005121203624967	true	3	16	1		86.43318176269531									N	N	N	N
1tunca7	actin	UNKNOWN	SYBR	None				27.6560001373291	27.43315315246582	0.2998186945915222					true	0.19068063569905777	true	3	22	1		90.45533752441406									N	N	N	N
1tunca7	actin	UNKNOWN	SYBR	None				27.091999053955078	27.43315315246582	0.2998186945915222					true	0.19068063569905777	true	3	21	1		90.45533752441406									N	N	N	N
1tunca7	actin	UNKNOWN	SYBR	None				27.551000595092773	27.43315315246582	0.2998186945915222					true	0.19068063569905777	true	3	21	1		90.45533752441406									N	N	N	N
1tungsk	actin	UNKNOWN	SYBR	None				28.69300079345703	28.691701889038086	0.09333206713199615					true	0.19068063569905777	true	3	22	1		90.25914001464844									N	N	N	N
1tungsk	actin	UNKNOWN	SYBR	None				28.78499984741211	28.691701889038086	0.09333206713199615					true	0.19068063569905777	true	3	23	1		90.25914001464844									N	N	N	N
1tungsk	actin	UNKNOWN	SYBR	None				28.597999572753906	28.691701889038086	0.09333206713199615					true	0.19068063569905777	true	3	22	1		90.25914001464844									N	N	N	N
1tungsk	trib3	UNKNOWN	SYBR	None	10.768038749694824	8.587308883666992	13.502561569213867	30.243999481201172	30.314367294311523	0.18102537095546722		1.6226673126220703	0.2036689966917038	-3.4286835193634033	true	0.19439522035735243	true	3	25	1		84.47115325927734									N	N	N	N
1tungsk	trib3	UNKNOWN	SYBR	None	10.768038749694824	8.587308883666992	13.502561569213867	30.179000854492188	30.314367294311523	0.18102537095546722		1.6226673126220703	0.2036689966917038	-3.4286835193634033	true	0.19439522035735243	true	3	25	1		84.47115325927734									N	N	N	N
1tungsk	trib3	UNKNOWN	SYBR	None	10.768038749694824	8.587308883666992	13.502561569213867	30.520000457763672	30.314367294311523	0.18102537095546722		1.6226673126220703	0.2036689966917038	-3.4286835193634033	true	0.19439522035735243	true	3	25	1		84.56925201416016									N	N	N	N

Replicate 3
3nt	hspa5	UNKNOWN	SYBR	None				20.770000457763672	20.781545639038086	0.013546253554522991					true	0.25257040819703624	true	3	15	1		86.62474060058594
3nt	hspa5	UNKNOWN	SYBR	None				20.797000885009766	20.781545639038086	0.013546253554522991					true	0.25257040819703624	true	3	15	1		86.52630615234375
3nt	hspa5	UNKNOWN	SYBR	None				20.777999877929688	20.781545639038086	0.013546253554522991					true	0.25257040819703624	true	3	15	1		86.42787170410156
3tun	hspa5	UNKNOWN	SYBR	None				18.47100067138672	18.503416061401367	0.03440316393971443					true	0.25257040819703624	true	3	13	1		86.52630615234375
3tun	hspa5	UNKNOWN	SYBR	None				18.5	18.503416061401367	0.03440316393971443					true	0.25257040819703624	true	3	13	1		86.42787170410156
3tun	hspa5	UNKNOWN	SYBR	None				18.538999557495117	18.503416061401367	0.03440316393971443					true	0.25257040819703624	true	3	13	1		86.42787170410156
3tunca7	hspa5	UNKNOWN	SYBR	None				21.406999588012695	21.429487228393555	0.02187357097864151					true	0.25257040819703624	true	3	16	1		86.52630615234375
3tunca7	hspa5	UNKNOWN	SYBR	None				21.450000762939453	21.429487228393555	0.02187357097864151					true	0.25257040819703624	true	3	16	1		86.42787170410156
3tunca7	hspa5	UNKNOWN	SYBR	None				21.43199920654297	21.429487228393555	0.02187357097864151					true	0.25257040819703624	true	3	16	1		86.42787170410156
3nt	trib3	UNKNOWN	SYBR	None				28.908000946044922	28.898958206176758	0.05202537402510643					true	0.23959995741232187	true	3	23	1		84.65608215332031
3nt	trib3	UNKNOWN	SYBR	None				28.945999145507812	28.898958206176758	0.05202537402510643					true	0.23959995741232187	true	3	23	1		84.55764770507812
3nt	trib3	UNKNOWN	SYBR	None				28.843000411987305	28.898958206176758	0.05202537402510643					true	0.23959995741232187	true	3	23	1		84.65608215332031
3tun	trib3	UNKNOWN	SYBR	None				24.687999725341797	24.702392578125	0.020757902413606644					true	0.23959995741232187	true	3	19	1		84.65608215332031
3tun	trib3	UNKNOWN	SYBR	None				24.691999435424805	24.702392578125	0.020757902413606644					true	0.23959995741232187	true	3	19	1		84.55764770507812
3tun	trib3	UNKNOWN	SYBR	None				24.72599983215332	24.702392578125	0.020757902413606644					true	0.23959995741232187	true	3	19	1		84.55764770507812
3tungsk	trib3	UNKNOWN	SYBR	None				27.847000122070312	27.783796310424805	0.06684623658657074					true	0.23959995741232187	true	3	22	1		84.65608215332031
3tungsk	trib3	UNKNOWN	SYBR	None				27.791000366210938	27.783796310424805	0.06684623658657074					true	0.23959995741232187	true	3	22	1		84.55764770507812
3tungsk	trib3	UNKNOWN	SYBR	None				27.714000701904297	27.783796310424805	0.06684623658657074					true	0.23959995741232187	true	3	22	1		84.45921325683594
3nt	actin	UNKNOWN	SYBR	None				24.10700035095215	24.12358856201172	0.11034904420375824					true	0.1807080435256173	true	3	18	1		90.5789794921875
3nt	actin	UNKNOWN	SYBR	None				24.240999221801758	24.12358856201172	0.11034904420375824					true	0.1807080435256173	true	3	19	1		90.38199615478516
3nt	actin	UNKNOWN	SYBR	None				24.02199935913086	24.12358856201172	0.11034904420375824					true	0.1807080435256173	true	3	18	1		90.38199615478516
3tun	actin	UNKNOWN	SYBR	None				25.871000289916992	25.817270278930664	0.09203243255615234					true	0.1807080435256173	true	3	20	1		90.38199615478516
3tun	actin	UNKNOWN	SYBR	None				25.8700008392334	25.817270278930664	0.09203243255615234					true	0.1807080435256173	true	3	20	1		90.28350830078125
3tun	actin	UNKNOWN	SYBR	None				25.711000442504883	25.817270278930664	0.09203243255615234					true	0.1807080435256173	true	3	20	1		90.28350830078125
3tunca7	actin	UNKNOWN	SYBR	None				24.83799934387207	24.863265991210938	0.027173949405550957					true	0.1807080435256173	true	3	19	1		90.28350830078125
3tunca7	actin	UNKNOWN	SYBR	None				24.892000198364258	24.863265991210938	0.027173949405550957					true	0.1807080435256173	true	3	19	1		90.18501281738281
3tunca7	actin	UNKNOWN	SYBR	None				24.858999252319336	24.863265991210938	0.027173949405550957					true	0.1807080435256173	true	3	19	1		90.18501281738281
3tungsk	actin	UNKNOWN	SYBR	None				24.371999740600586	24.45107078552246	0.0687810406088829					true	0.1807080435256173	true	3	19	1		90.38199615478516


Analysis:
For XBP1 splicing, all analyses was done with plate reader software. This is seen in the "Raw XBP1 data" tab.
Results displayed were transposed directly from these data. These results come from ddCT analysis. (unspliced XBP1 as the control)

For HSPA5 and Trib3, for replicates 1 and 2 calculations were initially done on the QPCR machine.
This provided relative quant values using the ddCT method (actin control)
As actin was run on a separate plate for the replicate 3 samples, ddCT was performed by hand. This is below:

Replicate 3 calculations:
	MEAN CT values				Relative quant (for NT)
	HSPA5	Trib3	Actin		HSPA5	Trib3	Actin
NT	20.782	28.899	24.124	NT	1	1

CA7	22.219		24.851	CA7	0.611320138846034
GSK		30.199	25.041	GSK		0.7668413298365465
NT Tun	18.503	24.702	25.817	NT Tun	15.692464063816837	59.30163596098375
								0.06792092890787867
Ca7 tun	21.429		24.863	Ca7 tun	1.0658467360679442
GSK tun		27.784	24.451	GSK tun		0.6164216886742478

For each replicate, HSPA5 and TRIB3 results were then normalized to the tunicamycin alone treatment condition results.
Pre normalization:
	1 NT	1 Tun	1 Tun CA7	1 Tun GSK	2 NT	2 Tun	2 Tun CA7	2 Tun GSK	3 NT	3 Tun	3 Tun CA7	3 Tun GSK
HSPA5	1	19.777463912963867	4.831404209136963		1	5.9979047775268555	2.0208442211151123		1	15.692464063816837	1.0658467360679442
Trib3	1	46.85881042480469		3.192003011703491	1	81.77969360351562		10.768038749694824	1	59.30163596098375		0.6164216886742478

Normalized inhibitor treatments:
	DMSO	DMSO	DMSO	CA7	CA7	CA7
HSPA5	1	1	1	0.24428835923548525	0.3369250256667757	0.06792092890787867
Trib3	1	1	1	0.06811959123089062	0.13167130219269882	0.010394682687671708
	Rep 1	Rep 2	Rep 3	Rep 1	Rep 2	Rep 3


	DMSO	DMSO	DMSO	DMSO	4u8c	4u8c	4u8c	4u8c
Spliced XBP1	145.838	204.159	236.307	231.719	0.052	0.036	0.038	0.041
(Relative Unspliced)

	(ALL tunicamycin treated)
	DMSO	DMSO	DMSO	CA7	CA7	CA7	GSK	GSK	GSK
HSPA5	1	1	1	0.24428835923548525	0.3369250256667757	0.06792092890787867
Trib3	1	1	1				0.06811959123089062	0.13167130219269882	0.010394682687671708
	Rep 1	Rep 2	Rep 3	Rep 1	Rep 2	Rep 3	Rep 1	Rep 2	Rep 3

BMDMs were derived from C57BL/6J mice and differentiated for a total of 8 days in DMEM+10% FBS + 20% L929 supernatant + sodium pyruvate + HEPES with a media addition 3 days post plating and a change of media 5 days post plating
Cells were then harvested and replated in DMEM + 10% FBS + HEPES + sodium pyruvate.
The following day, media was replaced with media containing inhibitors at 1x or received DMSO alone
2 hours post treatment, media was replaced with media containing tunicamycin and/or inhibitors (or dmso) as appropriate.
4 hours post treatment, media was removed from cells and TRIzol was added directly to wells.
RNA was then extracted from lysed samples using Direct-Zol RNA miniprep kit per manufacturers instructions (Zymo Research)
Reverse transcription was performed prior to qPCR using the iScript cDNA synthesis kit (Bio-Rad Laboratories)
Quantiative PCR was performed using Luna Universal qPCR Master Mix (New England BioLabs) using an Applied Biosystems Viia 7 Real-Time PCR system
ddCT method was used for calculating relative abundances.
The following primer sequences were used: spliced XBP1 F, 5’-ggtctgctgagtccgcagcagg-3’, R, 5’-gaaagggaggctggtaaggaac-3’ 57; unspliced XBP1 F, 5’-cagactatgtgcacctctgc-3’, R, 5’-cagggtccaacttgtccagaat-3’ 58; HSPA5 F, 5’- acttggggaccacctattcct-3’, R, 5’- atcgccaatcagacgctcc-3’ 59; Trib3 F, 5’- gcaaagcggctgatgtctg-3’, R, 5’-agagtcgtggaatgggtatctg-3’ 60–62; β-actin F, 5’- cgcagccactgtcgagtc-3’, R, 5’-ccttctgacccattcccacc-3’ 63.