Figure 8.
The failure of livers of Gank LKO mice to proliferate after CCl4injections is caused by high levels of tumor-suppressor proteins, which are eliminated by Gank in control LoxP mice. (A) Expression of TSPs in LoxP and Gank LKO mice after CCl4 injury. The experiment was performed with 3 biological replicates. (B) Calculations of levels of TSPs as ratios to β-actin using biological replicates. P values for HNF4α were as follows: 0 hours (P > .99), 24 hours (P > .99), 48 hours (P < .0002), 72 hours (P > .05), and 96 hours (P > .99). P values for Rb were as follows: 0 hours (P > .99), 24 hours (P > .18), 48 hours (P < .009), 72 hours (P > .33), and 96 hours (P > .99). P values for C/EBPα were as follows: 0 hours (P > .99), 24 hours (P > .19), 48 hours (P < .03), 72 hours (P < .009), and 96 hours (P > .86). P values for p53 were as follows: 0 hours (P < .002), 24 hours (P > .23), 48 hours (P < .02), 72 hours (P > .99), and 96 hours (P > .99). (C) Gank interacts with TSPs in livers of LoxP mice at 48 hours after CCl4 injury. Co-IP assay was applied for these studies. Nuclear extracts of LoxP and Gank LKO mice (200 μg for LoxP 0-hour time point and Gank LKO at 0 and 48 hours and 1 mg for LoxP at 48 hours) were incubated with antibodies to Gank and rabbit IgG beads (TrueBlot). After washing, IPs were loaded on the gel and probed with antibodies to Rb, p53, CUGBP1, C/EBPα, and HNF4α. The IPs were performed 3 times. Every IP was examined by Western blot with the earlier-mentioned antibodies. Right: Hypothetical mechanism for the failure of livers of Gank LKO mice to proliferate after CCl4-mediated injury. Two way analysis of variance. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001 LoxP vs Gank LKO.