Figure 1.

A pooled shRNA screen in BRAF-mutant human lung cancer cells identifies new modifiers of the RAF inhibitor response including YAP. (a) Summary of the pooled shRNA screening strategy in BRAF-mutant human lung cancer cells. (b,c) Primary screen data showing gene targets (b) and shRNAs depleted specifically upon vemurafenib treatment (c), highlighting YAP1 in red. shYAP1, shRNA to YAP1; shLUC, shRNA to the luciferase gene. (d) Validation of the effects of YAP1 knockdown on sensitivity to vemurafenib in HCC364 BRAF-mutant lung cancer cells (both IC₅₀ and cell viability results are shown). The inset shows the effects of each YAP1 shRNA by immunoblot for YAP protein expression. SCR, scrambled control shRNA. Data are shown as means ± s.e.m. (n = 3 biological replicates). (e) Validation of the effects of YAP1 knockdown on sensitivity to trametinib in HCC364 BRAF-mutant lung cancer cells (IC₅₀, cell viability and maximal growth inhibition results are shown). Data are shown as means ± s.e.m. (n = 3 biological replicates). (f) Effects of YAP1 knockdown on sensitivity to vemurafenib and trametinib in HCC364 BRAF-mutant lung cancer cells (cell growth by crystal violet staining assays is shown, with quantification for each condition relative to cells expressing the scrambled control shRNA treated with DMSO control). (g) Effects of YAP1 knockdown on sensitivity to trametinib in Cal-12T BRAF-mutant (non-V600E) lung cancer cells (IC₅₀, cell viability and maximal growth inhibition results are shown). Data are shown as means ± s.e.m. (n = 3 biological replicates).