Fig. 5

Effect of EGF and/or cetuximab treatment on EGFR, MAPK and AKT activation in Hs746T cells. Phosphorylation of EGFR was detected in total cell lysates by Western blot analysis using antibodies directed against a pEGFR (Y1068), b pMAPK (T202/Y204) and c pAKT (S473). α-tubulin was used as a loading control. The depicted results are representative of three independent experiments. Each membrane was probed with antibodies specific for a pEGFR (Y1068), c pAKT (S473) and α-tubulin (loading control). Each blot was then stripped and re-probed with an antibody specific for b pMAPK (T202/Y204). One representative Western blot analysis for each stimulation period is shown. The average phosphorylation levels were quantified using densitometric analysis and were calculated in relation to the levels of α-tubulin (+SD). Statistical p-values are listed in Additional file 1: Table S6. Additionally, one representative Western blot of each stimulation period is shown. Abbreviation: Cet = cetuximab