Figure 2.

Knockdown of Arid1b alleviates OIS in vivo and cooperates in HCC formation. (A) Scheme of the in vivo senescence experiments. Transposon constructs carrying Nras^G12V cDNA and Arid1b shRNAs (or control) were delivered into mouse livers by hydrodynamic injection (Nras^G12V/D38A was used as negative control). n = 4 mice per condition were used. (B) Quantification of Arid1b nuclear IF staining in Nras⁺ mouse hepatocytes in the above-described conditions is shown. (C) Representative pictures of immunohistochemistry (IHC) for Nras, SA-β-Gal staining, and IF staining for p21^CIP1 and p16^INK4a on liver sections from mice injected with the indicated vectors. (D–F) Quantification of the SA-β-Gal staining and percentages of Nras-positive, p16^INK4a-positive, or p21^CIP1-positive cells (D) and 53bp1-positive (E) or Ki67-positive (F) cells in liver sections from mice injected with the indicated vectors. Graphs represent mean ± SD from n = 4 mice. (***) P < 0.001; (**) P < 0.01; (*) P < 0.05; (n.s.) nonsignificant by two-tailed Student t-test. (G, left) Scheme of the in vivo tumor growth experiment. Transposon constructs carrying Nras^G12V cDNA and an Arid1b shRNA (or control) were delivered into mouse livers by hydrodynamic injection. Mice were monitored for 30 wk. n = 10 shCTR and n = 13 shArid1b.1 mice were used. Kaplan-Meier survival curves (middle) and representative pictures of livers (right) showing GFP-positive tumor nodules as they were derived from the injected constructs. (*) P < 0.05 by log rank (Mantel-Cox) test. Bar, 100 μm. DAPI was used to visualize the nuclei.