Fig. 5: JMJD3 coactivates PPARα to induce hepatic autophagy.

a Venn diagram (top) for genes with PPARα cistrome detected by ChIP-seq and hepatic genes inhibited by liver JMJD3 downregulation (as shown in Fig. 1b). G/O analysis (bottom) of the overlapping genes. b WT or PPARα-KO mice were fasted (Fs) for 24 h or refed for 24 h (Fd) after fasting. Levels of LC3 in liver extracts measured by IB with the LC3-II/I ratios shown below the blot (top, n = 3 mice). The mRNA levels of indicated genes (bottom, n = 5 mice). c PMH from JMJD3-floxed mice were infected with AAV-GFP or AAV-Cre for 72 h and treated with vehicle or WY14643 for 12 h. The indicated proteins were detected by IB with the LC3-II/I ratios shown below the blot (top, n = 3 culture dishes). The mRNA levels of indicated genes (bottom, n = 5 mice). d C57BL/6 or PPARα-KO mice were fasted for 1 h and treated with vehicle or 0.1 mg/kg FGF21 for 3 h. Hepatic levels of LC3 measured by IB with the LC3-II/I ratios shown below the blot (top, n = 3). The mRNA levels of indicated genes (bottom, n = 5 mice). e LC3 and p62 detected by IHC. Representative images of liver sections (left) and the average number of puncta/cell (right, n = 10 hepatocytes) are shown (scale bar = 10 μm for LC3, 50 μm for p62). f re-ChIP: Hepatocytes were transfected with PPARα siRNA, 72 h later, cells were treated with FGF21 for 2 h. PPARα was immunoprecipitated followed by immunoprecipitation of JMJD3, and enrichment of Tfeb, Atg7, and Atgl sequences was determined (n = 3 culture dishes). g Hepa1c1c7 cells were transfected with a luciferase reporter containing the PPARα binding site or mutated site from Tfeb or Atg7 and with plasmids and siRNAs as indicated. After 36 h, the cells were treated with 50 μM WY14643 and 100 ng/ml FGF21 overnight. Luciferase activities were normalized to β-galactosidase activities (n = 4 culture dishes). Source data are provided as a Source Data file. b–g Values are presented as mean ± SD. Statistical significance was measured using the g one- or b–f two-way ANOVA with the Bonferroni post-test. **P < 0.01.