Fig. 7
Response of BRAFV600E mutant MaMel cells to PI3K/AKT pathway inhibition. A Representative Western blot analysis of AKT and S6 phosphorylation in MaMel cells treated with dactolisib (BEZ), vemurafenib (PLX), or trametinib (TR). The phosphorylated (p-) and total proteins are shown; GAPDH served as loading control. 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 five independent experiments are shown (n = 5). C Fold change reduction of the cell proliferation rate (−log2 FC) of MaMel cells under inhibition with BEZ at the indicated time points. Data from three independent experiments are shown (n = 3). The time points outside the brackets indicate the total time of cells in culture. Time points in brackets indicate the inhibition time. D Representative side population (SP) assay detecting the efflux of DyeCycle Violet stain (DCV) in MaMel21 under inhibition for 66 h with PLX, TR, BEZ, or the ABCG2 inhibitor Ko143. The polygonal gates indicate the SP subsets (DCV-) in each condition. E Quantification of the percentage of SP in MaMel21 under inhibition. Data from five independent experiments are shown (n = 5). All bar plots show the mean ± SD of the individual experiments. #p < 0.1, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001