Figure 1
PLX4720 inhibits proliferation and induces apoptosis in mutant B-RAF melanoma cells. (a) Whole cell lysates from Mel-RM (wild-type B-RAF) and Mel-RMu (B-RAFV600E) cells treated with PLX4720 at indicated concentrations for 72 h (upper panel) or at 10 μM for indicated periods (lower panel), were subjected to western blot analysis of phosphorylated ERK1/2 and ERK1/2. (b) Mel-RM and Mel-RMu cells were treated with PLX4720 at indicated concentrations for 72 h. Cell viability (left panel) and apoptosis (right panel) were quantitated by the MTS assay and propidium iodide (PI) method, respectively. The data shown are the mean±S.E. of three individual experiments. (c) Mel-RM and Mel-RMu cells were treated with PLX4720 at 10 μM for indicated periods. Cell viability (left panel) and apoptosis (right panel) were quantitated by the MTS assay and PI method, respectively. The data shown are the mean±S.E. of three individual experiments. (d) A summary of the effect of PLX4720 on cell survival in a panel of mutant and wild-type B-RAF melanoma cell lines. Cells treated with PLX4720 at 10 μM for 72 h were subjected to MTS assays. The data shown are the mean±S.E. of three individual experiments. (e) Left panel: B-RAFV600E Mel-RMu and Mel-CV cells were transfected with the control or B-RAF siRNA. After 24 h, whole cell lysates were subjected to western blot analysis of B-RAF, phosphorylated ERK1/2, and ERK1/2. Western blot analysis of A-RAF and C-RAF was included as controls to show the specificity of the B-RAF siRNA. Right panel: Mel-RMu and Mel-CV cells were transfected with the control or B-RAF siRNA. After 24 h, cells were treated with PLX4720 (10 μM) for a further 72 h. Apoptotic cells were measured by the PI method. The data shown are either representative (left panel), or the mean±S.E. (right panel), of three individual experiments
