Figure 6.

Verification of results in independent BRAF knockout cells. A: Cell death was assessed by determining the sub G1-fraction by flow cytometry and analyzed on a molecular level by Western blotting for PUMA. B-D: In chemosensitivity assays RAF265 and vemurafenib did not induce different responses, while with dabrafenib, the wild type clone showed a 2.6 fold higher IC₅₀ value as compared to control clones. E-F: In Western blotting experiments and subsequent densitometry analysis, IC₅₀ concentrations of dabrafenib showed a stronger effect on relative phosphorylation levels of Mek 1/2 and Erk 1/2 in parental and BRAF-mutant cells than RAF265 and vemurafenib. G: In proliferation assays, no differential responses were observed for PI-103. H: PIK3CA status was confirmed by sequencing. I: Phosphorylation of AKT was examined by Western blotting.