Fig. 5: Increased proliferation of wild-type cells is sufficient to counteract expansion of HrasG12V/+ cells in mosaic skin.
From: Injury prevents Ras mutant cell expansion in mosaic skin
a, Violin plots showing cell scoring based on expression of EGFR ligands (Extended Data Fig. 6i), separated by cell type and genotype. Two-tailed t-test comparing the averages of biological replicates according to conditions (fibroblasts: n = 3 wild-type mosaic and n = 6 HrasG12V/+ mosaic; epithelial cells n = 6 mice per genotype), P values are shown. Internal box plots denote the 25th, 50th and 75th centiles, with whiskers depicting minima and maxima, excluding outliers that are beyond 1.5× the interquartile range. b, Western blot analysis (left) and quantification (middle) of phosphorylated EGFR (p-EGFR) and total EGFR in injured and uninjured conditions (n = 3 mice). Paired, two-tailed t-test. Pairs of coloured dots represent ratios for individual mice. Right, western blot analysis of total EGFR normalized to GAPDH (n = 3 mice). Blots were processed at the same time. Unpaired, two-tailed t-test. c, Quantification of mitotic cells in tdTomato+ and tdTomato− areas. d, Left, representative two-photon revisit images following injury. Right, the increase in tdTomato+ area after tamoxifen induction. In c,d, vehicle, n = 3 mice; Gefitinib, n = 4 mice. e, Quantification of mitotic cells in tdTomato+ and tdTomato− areas in wild-type mosaic (n = 3 mice), HrasG12V/+ mosaic (n = 3 mice) and constitutive p21-null HrasG12V/+ mosaic (n = 4 mice). f, Quantification of p-histone H3-positive cells in tdTomato+ and tdTomato− areas (n = 6 mice). g, Representative two-photon revisit images following injury in wild-type mosaic, HrasG12V/+ mosaic and constitutive p21null-HrasG12V/+ mosaic mice. h, The increase in tdTomato+ area. Unpaired, ordinary one-way ANOVA comparing wild-type mosaic, HrasG12V/+ mosaic and constitutive p21null-HrasG12V/+ mosaic at different time points in the uninjured condition. In g,h, n = 3 wild-type mosaic mice, n = 3 HrasG12V/+ mosaic mice and n = 4 constitutive p21null-HrasG12V/+ mosaic mice. c–f, Unpaired or paired two-tailed t-test for comparisons between different groups or within the same group of mice. P values are shown. At least three independent areas of approximately 300 µm2 were analysed for each mouse (Methods). Data are mean ± s.d.
