Detailed analysis of post‐translational compensation of varying Erk concentration. Detailed analysis of post‐translational compensation of varying Erk.

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Detailed analysis of post‐translational compensation of varying Erk concentration. Detailed analysis of post‐translational compensation of varying Erk concentration. (A) Position of mutations of the analysed cells in the pathway: five colon carcinoma cell lines were analysed, LIM1215 has no mutation in the MAPK signalling pathway, HT29 and RKO express constitutively active B‐Raf (V600E), and SW480 and HCT116 harbour an activating mutation in K‐Ras. (B) Changes in expression of Erk1 (left) and Erk2 (right) 48 h after treating the cells with scrambled control siRNA or siRNA targeting Erk1, Erk2 or both isoforms in the five cell lines. If one isoform is knocked down, no significant change in the other isoform can be observed. (C) Pan‐isoform Erk and phospho‐Erk levels after knockdown of Erk1 and/or Erk2 were calculated as fraction of the unperturbed scrambled controls. Cells with B‐Raf mutation show a linear relation between Erk concentration and phospho‐Erk level that is predicted by a mathematical model for a system without feedback (shown as line). Cells with B‐Raf wild type show strong robustness in phospho‐Erk level corresponding to response coefficients of 0.36 and 0.20 for HCT116 and SW480, respectively. (D) Representative western blot images of knockdown experiments in SW480 and HT29. (E) Changes in Mek phosphorylation 48 h after treating the cells with scrambled control siRNA or siRNA against Erk1, Erk2 or both isoforms. While B‐Raf wild‐type cells show a strong increase in phospho‐Mek, B‐Raf‐mutated cells (HT29 and RKO) show no change in phospho‐Mek after knockdown. Source data is available for this figure at www.nature.com/msb. Raphaela Fritsche‐Guenther et al. Mol Syst Biol 2011;7:489 © as stated in the article, figure or figure legend