Fig. 2

Effect of Itga6 deletion on growth and differentiation of Brca1/p53-deficient tumors. A Immunofluorescent staining with anti-Itgα6 (white), and anti-Ki67 (red) antibodies. The graph shows the percentage of Ki67 + cells (mean ± SEM) in small (< 750 mm³; n = 8 animals per genotype) and large (> 750 mm³; n = 7 animals per genotype) tumors. B Immunofluorescent staining with anti-K8 (white), and anti-cleaved caspase 3 (CC3, red) antibodies. The graph shows the percentage of CC3 + cells (mean ± SEM) in small (< 750 mm³; n = 4 animals per genotype) and large (≥ 750 mm³; n = 4 animals per genotype) tumors. A and B: Nuclear DAPI staining is shown in blue. Scale bar: 20 µm. C Tumor growth curves of Brca1p53-KO (n = 7) and α6KO/Brca1p53-KO (n = 7) tumors. The difference between groups is not significant (Kruskal–Wallis test). D Upper panels: Representative FACS analysis of a Brca1p53-KO and a α6KO/Brca1p53-KO tumor with CD24 and Lin (CD31/CD45/Ter-119) antibodies. The red rectangle shows the CD24⁺ Lin⁻ population sorted for gene expression analysis. Lower panels: Representative FACS analysis of tumors with CD24 and CD49f (α6-integrin) antibodies showing depletion of surface Itgα6 in most epithelial cells of the α6KO/Brca1p53-KO tumor. E RT-qPCR analysis of the CD24⁺ Lin⁻ tumor cell population from Brca1p53-KO (n = 5) and α6KO/Brca1p53-KO (n = 5) animals (mean ± SEM). F Representative immunofluorescent staining with anti-K5 (red) and anti-K8 (white) antibodies. Magnification of the white rectangle is shown in the right panels of Brca1p53-KO and α6KO/Brca1p53-KO tumors. Nuclear DAPI staining is shown in blue. Scale bar: 75 µm (left panels for each tumor) and 20 µm (magnification in the right panels). In A, B, E *P < 0.05, n.s., non-significant