p53 inhibits migration and invasion of E1A;Hras^G12V;H11^Cas9 MEFs. (A) Representative bright-field images of sgRNA-targeted E1A;Hras^G12V MEFs migrating through Boyden chamber filter and stained with Giemsa. Scale bar, 100 µm. (B) Mean fold change in migrating E1A;Hras^G12V MEFs ± SD, expressed relative to counterpart sgNTC control cell line. n = 3 cell lines/sgRNA; *, P < 0.05 **, P < 0.005, one-way ANOVA, Bonferroni’s multiple comparisons test. (C and E) Representative images of sgNTC and sgp53(2) E1A;Hras^G12V;H11^Cas9 cultured in a 3D-collagen matrix in 5% (C) or 21% (E) O₂. WGA marks cell membranes, and DAPI marks nuclei. Scale bar, 0.2 mm. (D) Percentage invading colonies in 5% O₂. Data are mean ± SD, n = 3 cell lines/sgRNA; ***, P < 0.0001, one-way ANOVA, Bonferroni’s multiple comparisons test. (F) Percentage invading colonies in 21% O₂. Data are mean ± SD, n = 3 cell lines/sgRNA; ns, not significant (P > 0.05), Mann–Whitney unpaired t test.