Fig. 5: Neddylation deficiency autonomously activates hepatocyte fetal reprogramming.
a Representative Western blot and relative mRNA levels of mature hepatocyte and BEC/ progenitor-specific marker genes in mouse Nae1f/f primary hepatocytes (PH) infected with pAd-GFP and pAd-Cre for 48 h (n = 6 per group). pAd-GFP (black), pAd-Cre (red). Multiple unpaired t-tests with the Holm-Sidak method. b HepG2 cells were infected with pLentiCRISPR/Cas9 V2 lentiviruses bearing two specific human NAE1 sgRNAs to generate bulk cultures with NAE1 knockout (KO1 and KO2), respectively. Cells infected with viruses expressing no sgRNA were used as control (V). Western blot and relative mRNA levels of mature hepatocyte and BEC/progenitor-specific marker genes were analyzed (n = 4 per group). V (black), KO1 (red), KO2 (blue). Two-way ANOVA followed by Tukey’s multiple comparisons test. c Representative immunofluorescence staining for KRT19 and quantification of KRT19+ cells (n = 5 per group). Scale bar: 100 μm. V (black), KO1 (red), KO2 (blue). One-way ANOVA with Tukey’s multiple comparisons test. d Representative Western blot and relative mRNA levels of mature hepatocyte and progenitor/BEC-specific marker genes in HepG2 cells treated with vehicle (Veh) or indicated concentrations of MLN for 48 h (n = 3 per group). Veh (black), MLN0.2 (red), MLN2.0 (blue). Two-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.005. All quantitative data were presented as mean ± SEM. Source data are provided as a Source Data file.
