Fig. 2: JMJD3 promotes hepatic autophagy, including lipophagy.

a The indicated hepatic proteins were detected by IB in JMJD3-floxed mice infected with AAV-TBG-Cre or AAV-GFP for 12 weeks and fasted for 16 h. Relative band intensities for p62 and the LC3-II/LC3-I ratios are below the blots (left, n = 5 mice). LC3 and p62 in representative images of liver sections detected by IHC (middle) and numbers of LC3 puncta/cell (right, n = 10 hepatocytes) (scale bar = 10 μm for LC3, 50 μm for p62). b Levels of the indicated hepatic proteins were determined by IB in C57BL/6 mice infected with Ad-empty (GFP) or Ad-JMJD3 for 4 weeks and fasted for 8 h (n = 3 mice). c, d Hepa1c1c7 cells were transfected with expression plasmids or JMJD3 siRNA as indicated for 72 h and cultured in HBSS for 2 h. c Representative confocal images and the average number of GFP-LC3-II puncta/cell (right, n = 10 cells) are shown (scale bar = 5 μm). d Cells were stained for lipid droplets with BODIPY (red) and imaged by confocal microscopy. Co-localization of GFP-LC3 puncta and BODIPY in the merged image is indicated by white arrows. The number of fluorescent puncta that co-localized with lipid staining (right, n = 20 cells) are shown (scale bar = 5 μm). e–g C57BL/6 mice were fed a normal chow diet (ND) or high-fat diet (HFD) for 8 weeks and then were injected with adenoviruses as indicated for 4 weeks. e Hepatic levels of the indicated proteins were measured by IB (n = 3 mice). The ratios of band intensities for the LC3-I/LC3-II relative to those in the first lane are shown below the blot. f Lipids in liver sections stained with Oil Red O (ORO) and H&E (scale bar = 50 μm). g Levels of hepatic triglycerides (TG) and serum β-hydroxybutyrate (β-HDB) (n = 5 mice). Source data are provided as a Source Data file. All values are presented as mean ± SD. Statistical significance was measured using the (a, c, d) Mann–Whitney test or g two-way ANOVA with the Bonferroni post-test. *P < 0.05, **P < 0.01, and NS statistically not significant.