Fig. 6

Hypermorphic BRAF variants confer resistance to genotoxic stress in LUAD. a Association between integral survival and BRAF genotype in 28 LUAD cell lines. Red bar represents mutation. BRAF mutation ranked 12 out of 6743 genomic features after outlier exclusion. The P-values were calculated using the empirical permutation test. b Immunoblot analysis of representative cell lines profiled for radiation sensitivity. c BEAS-2B cells stably infected with vector alone (ϕ) or vector expressing BRAF variants were profiled for RAF-MEK-ERK pathway activity by immunoblot. d BEAS-2B cells in c were treated with ionizing radiation and cell number was determined on days 7–9. Representative curves are shown. Data points represent mean ± s.e.m. The heatmap of integral survival is organized by the order of all of the transduced cells in c. e Schematic depiction of the experimental design used in f. f BEAS-2B cells expressing BRAF G466V or wild-type (WT) were injected into the flank of NSG mice and block randomized into each treatment arm: Mock (ϕ) or X-ray (2 Gy × 3). Tumor volumes were measured at least twice weekly. Data represent the mean. Solid line represents the interpolation of mean using a third order polynomial fit. Dashed lines represent the 95% confidence interval of the polynomial fit. n = 5 independent animals for each condition. g The ratio of G466V to WT was determined in the harvested tumors (at a volume of ~500 mm³) using ddPCR. The proportion in each arm was normalized to the fractional abundance in cells expressing G466V alone. Data are expressed as the mean ± s.e.m. of three independent experiments