Figure 3.

Rank overexpression induces senescence through p16/p19 (Ink4a/Arf) in MECs and MEFs

(A) Representative images and quantification of SA-β-gal staining in MECs isolated from WT MGs and WT and p16/p19^−/− MEFs 6 days after infection with Rank or RASV12 overexpressing and control (ϕ) vectors. Number of independent experiments is shown. Quantifications were performed in triplicates and each dot represents a replica.

(B) Representative images and quantifications of Ki67⁺ (green) and p19⁺ (red) cells within total nuclei (DAPI, blue) in WT MECs and MEFs 6 days after infection with Rank-overexpressing and control (ϕ) lentivirus.

(C) Representative images and quantification of the H&E with SA-β-gal staining in MGs of the indicated genotypes. Three representative pictures of the indicated number of MGs were quantified.

(D) Representative images and quantification of SA-β-gal staining in Rank^+/tg and Rank^+/tg p16/p19^−/− MECs (ex vivo) cultured for 8 days. Quantifications were performed in triplicates and each dot represents a replica of three independent MGs/mice.

(E) Quantification of γH2AX+ foci per nuclei in the indicated cells 6 days after infection with control (ϕ) and Rank-overexpressing virus.

(F) Quantification of γH2AX+ cells relative to the epithelial area in the MGs of WT, Rank^+/tg, p16/p19^−/−, and Rank^+/tg p16/p19^−/− females. The number of mice analyzed is indicated for each genotype. Representative images are shown in Figure S3A. In all panels mean, SEM and t test p values are shown. See also Figures S2 and S3.