Fig. 3
Response of BRAFV600E mutant MaMel cells to MAPK/ERK pathway inhibition. A Representative Western blot analysis of MEK and ERK phosphorylation in MaMel cells treated with vemurafenib (3 µM, PLX; upper panel) or trametinib (5 nM, TR; lower panel). The phosphorylated (p-) and total proteins are shown; GAPDH served as loading controls. B Densitometric analysis of Western blots showing the fold change reduction (−log2 FC) of p-MEK and p-ERK after treatment with PLX or TR, in comparison to the corresponding vehicle (DMSO)-treated cells. Data from six independent experiments are shown (n = 6). C Quantification of the apoptotic response of MaMel cells under inhibition with PLX (3 µM) or TR (5 nM). Annexin V and PI co-staining, detected by flow cytometry, was used to define viable and early and late apoptotic cell populations at indicated time points. Data from three independent experiments are shown (n = 3). Time points outside the brackets indicate the total time of cells in culture; time points in brackets indicate the inhibition time. All bar and dot plots show the mean ± SD of the individual experiments. #p < 0.1, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001