BRAFV600E is required and sufficient to suppress AKT phosphorylation in a subset of melanoma cells.

(A) Western blot analysis of AKT phosphorylation 24 hours after BRAF or CRAF knock-down in A375 melanoma cell line 4 hour post RG7167 treatment. (B) Western blot analysis of AKT substrates (FOXO1, GSK3α/β, PRAS40) phosphorylation 24 hours after BRAF knock-down in A375 melanoma cell line. (C) Western blot analysis of AKT phosphorylation 24 hours after BRAF knock-down in LOX melanoma cell line. (D) Western blot analysis of AKT, MEK, and ERK phosphorylation in CHL1 melanoma cell line 24 hours after transient transfection (wild-type BRAF or BRAFV600E) and treated with RG7167 for 4 hours.