Figure 4.

Enforced expression of B-RAF^V600E inhibited Bim_S expression in wild-type B-RAF melanoma cells and melanocytes. (a) Mel-RM and Me1007 cells were stably transfected with cDNA encoding B-RAF carrying the V600E mutation. Whole cell lysates were subjected to western blot analysis of B-RAF, pERK1/2, and ERK1/2. The data shown are representative of three individual western blot analyses. (b) Mel-RM and Me1007 cells were stably transfected with cDNA encoding B-RAF carrying the V600E mutation. Cells were treated with SBHA (10 μg/ml) for a further 16 h. Total RNA was isolated and subjected to real-time PCR analysis for Bim_EL, Bim_L, and Bim_S mRNA expression. The levels of the expression of individual species before treatment were arbitrarily designated as 1. The data shown are the mean±S.E. of three individual experiments. (c) Whole cell lysates from Mel-RM and Me1007 cells treated as in b were subjected to western blot analysis of Bim and GAPDH (as a loading control). The arrowhead points to nonspecific bands. The data shown are representative of three individual western blot analyses. (d) Melanocytes were transiently transfected with cDNA encoding B-RAF carrying the V600E mutation. After 24 h, whole cell lysates were subjected to western blot analysis of B-RAF, pERK1/2, ERK1/2, Bim, and Bcl-2. The arrowhead points to nonspecific bands. The data shown are representative of three individual western blot analyses