Figure 5.

FAM150A-Activated ALK Phosphorylation in Melanoma Cells and Stimulated BRAFi Resistance

(A) A375 parental cells were cultured with conditioned medium harvested from A375 parental (A375-P CM) or vemurafenib-resistant A375 cells (A375-R CM), then treated with DMSO or the indicated concentrations of vemurafenib for 3 days, and analyzed for survival using the MTT assay. Relative cell survival (%) relative to DMSO-treated cells is shown.

(B) mRNA expression for FAM150A was measured for the indicated pairs of parental and BRAFi-resistant cells (A375, SKMEL-239, M229). mRNA expression is shown relative to respective parental melanoma cells. ACTINB mRNA expression was used for normalization.

(C) mRNA expression for FAM150A was measured in A375 cells treated with DMSO or vemurafenib (1 μM) for 24 h FAM150A mRNA expression relative to DMSO-treated A375 cells is shown. ACTINB mRNA expression was used for normalization.

(D) A375 cells overexpressing empty vector (−) or FAM150A (+) were analyzed for indicated proteins by immunoblotting. ACTINB was used as a loading control.

(E) A375 cells ectopically expressing an empty vector (EV) or FAM150A were treated with DMSO or the indicated concentrations of vemurafenib for 3 days and analyzed for survival by the MTT assay. Relative cell survival (%) relative to DMSO-treated cells is shown.

Data are presented as mean ± SD. *p < 0.05, ***p < 0.001, ****p < 0.0001. See also Figure S4.