FIGURE 5.

Depletion of PKN1 increases apoptosis initiated by a BRAF-V600E inhibitor. A375 and Mel-624 malignant melanoma cells were transfected with the indicated siRNA oligonucleotides. 48 h later, the cells were treated with BRAFi (2 μm PLX-4720 or as indicated) and medium conditioned with WNT3A-secreting (W3ACM) or vehicle-secreting (LCM) L cells. A, reporter activity was assayed in A375 malignant melanoma cells stably expressing a β-catenin reporter driving expression of firefly luciferase and a constitutive promoter driving Renilla luciferase. Flow cytometry was used to assay the percentage of A375 (B and C) and Mel-624 (D) cells expressing AnnexinV. Representative plots (B) and quantification of three independent experiments (C and D) are presented. Error bars represent the S.D. from at least three independent replicates, and the p values (**, p < 0.01; ***, p < 0.001) were calculated using a two-way ANOVA followed by a Bonferroni post-test. CTRL, control.