Fig. 3

Fig. 3. In the absence of Irf2bp2a, p53 pathway is aberrantly activated due to impaired turnover, which contributes to the defective liver development. (A) WISH assay of lfabp at 72 hpf in irf2bp2a^−/− embryos, and irf2bp2a^−/− embryos injected with p53 MO. (C) WISH assay of lfabp in WT, WT embryos injected with irf2bp2a MO, p53^−/−, and p53^−/− embryos injected with irf2bp2a MO. (B, D) The relative liver area measured. The data shown is fold difference compared with the level detected in control embryos. p values are denoted by asterisks. (Student t-test, N ≥ 3. Error bars represent mean ± SD, ns, not significant, ****P < 0.0001). (E) IRF2BP2 and P53 mRNA levels were measured by RT-qPCR in HepG2 cells after transfection of an irrelevant shRNA (NC, non-specific control) or a shRNA against IRF2BP2 (sh-IRF2BP2). (Student t-test, N ≥ 3. Error bars represent mean ± SD, ns, not significant, ***P < 0.001). (F) Western blot analysis of IRF2BP2, P53, IRF2 and MDM2 in WT and IRF2BP2-knockdown HepG2 cells. (G) IRF2 protein was immunoprecipitated (IP) with an anti-IRF2 antibody in WT and IRF2BP2-knockdown HepG2 cells, then MDM2 protein was detected by western blot analysis. (H) WISH assay of lfabp in WT, WT embryos injected with full length irf2 or irf2bp2a mRNA.