Figure 2: Model calibration with time-resolved quantitative immunoblot data of mCFU-E cells and BaF3-EpoR cells.

Growth-factor deprived mCFU-E cells (5x106 cells per condition) and BaF3-EpoR cells (1x107 cells per condition) were stimulated with different Epo doses and absolute concentrations were determined for pEpoR (B), pAKT (C), ppERK (D). The scale for pS6 (E) was estimated in arbitrary units. GTP-Ras (F) and ppERK were determined upon stimulation with indicated, color-coded Epo doses. pEpoR was analyzed by immunoprecipitation followed by immunoblotting, GTP-Ras was analyzed after pulldown using a fusion protein harboring GST fused to the Ras binding domain of Raf-1 followed by detection by quantitative immunoblotting. For pAkt and ppERK, cellular lysates were subjected to quantitative immunoblotting. Calibrator proteins were used for EpoR, AKT, GTP- Ras and ERK to facilitate the conversion to nM concentrations.

SED-ML simulations Fig. 2B - 2E: https://jjj.bio.vu.nl/models/experiments/adlung2017_fig2bto2e/simulate Fig. 2F: https://jjj.bio.vu.nl/models/experiments/adlung2017_fig2f/simulate Fig. 2G: https://jjj.bio.vu.nl/models/experiments/adlung2017_fig2g/simulate