Fig. 4: FGF21-induced hepatic autophagy is largely dependent on JMJD3.

a–c JMJD3-floxed mice were infected with AAV-TBG-Cre or AAV-GFP for 12 weeks (n = 6 mice/group), and injected i.v. with vehicle or FGF21 (0.1 mg/kg) for 3 h. a The indicated hepatic proteins were detected by IB (n = 3 mice). b LC3 or p62 was detected in liver sections by IHC and the number of LC3-II puncta/cell were quantified (right, n = 10 hepatocytes) (scale bar = 10 μm for LC3, 50 μm for p62). c Hepatic mRNA levels of the indicated autophagy-related genes measured by q-RTPCR (n = 5 mice). d, e PMH from JMJD3-floxed mice were infected with AAV-TBG-GFP or AAV-TBG-Cre for 72 h, and treated with vehicle or FGF21 (100 ng/ml) for 12 h. d Levels of the indicated proteins measured by IB (n = 3 culture dishes). e Levels of cellular triglycerides (TG) (n = 10 culture dishes). f Hepa1c1c7 cells were transfected with GFP-LC3 plasmid and with control (siC) or JMJD3 siRNA as indicated. After 72 h, cells were supplemented with 400 μM oleic acid for 6 h before incubation in serum-free DMEM containing vehicle or 100 ng/ml FGF21 for 12 h. Cells were stained for lipid droplets with BODIPY (red) and imaged by confocal microscopy. Co-localization of GFP-LC3 puncta (green) and BODIPY in the merged images is indicated by white arrows. The average number/cell (right, n = 20 cells) of fluorescent puncta that co-localized with lipid staining is shown (scale bar = 5 μm). Source data are provided as a Source Data file. All values are presented as mean ± SD. Statistical significance was measured using the (b, c, e, f) two-way ANOVA with the Bonferroni post-test. **P < 0.01, and NS statistically not significant.