Skip to main content
. 2019 Aug 8;33(1):74–85. doi: 10.1111/pcmr.12812

Figure 4.

Figure 4

RANKL diminishes BRAF inhibitor activity via MITF‐induced survival signalling (a). Growth curves of indicated cell lines with or without BRAFi (vemurafenib 1 µM WM98, 0.5 µM A375) in the presence or absence of RANKL (50 ng). Confluency was measured by time‐lapse microscopy using an Incucyte system (Mean ± SD, n = 3) (b). Protein expression of phospho‐ERK, MITF, beta tubulin (BTUB) and ERK, and quantification of relative cell number as determined by crystal violet staining of A375 cells. Cells were treated for 72 hr with/without 50ng/ml RANKL, the final 48 hr cells were treated with either DMSO or 0.5 µM vemurafenib in the presence of either a Scrambled control or a MITF specific siRNA (20 nM). (Mean ± SEM, n = 3) (c). RT‐qPCR analysis of gene expression in indicated melanoma cell lines treated for 24 hr with/without 50ng/ml of RANKL. (Mean ± SEM, n = 4) (d). Protein expression of BCL2 and ERK in A375 cells with/without 50ng/ml of RANKL at indicated time points. Relative BCL2 expression was quantified using Image J and normalized to ERK (e). Protein expression of MITF, ERK and RT‐qPCR analysis of gene expression in indicated melanoma cell lines, treated for 24 hr with/without 50ng/ml of RANKL, following previous 24 hr transfection with indicated siRNA. (Mean ± SEM, n = 3)