Loss of STARD4 can eliminate the protective effect of GPR35 against steatohepatitis (n = 6). Gpr35hep-NC2 and Gpr35hep-OE mice were injected (i.v.) with AAV8-TBG-Mir30-m-shRNA(NC)-mCherry or AAV8-TBG-Mir30-m-shRNA(Stard4)-mCherry to generate control (Stard4hep-NC2-Gpr35hep-NC2 and Stard4hep-NC2-Gpr35hep-OE) mice and hepatocyte STARD4-knockout (Stard4hep-KD-Gpr35hep-NC2 and Stard4hep-KD-Gpr35hep-OE) mice, which were then fed a HFCF diet for 16 weeks. (A) TC, FC, CE and (B) bile acid levels in the liver. (C) Representative Western blot of CYP7A1, CYP8B1, CYP27A1, CYP7B1, SR-BI, and ACAT2 proteins in the liver. (D) Body weight was measured every 4 weeks from 0 to 16 weeks. (E) Liver weight. (F) TG and (G) FFA levels in the liver. (H) H&E staining of liver tissue sections. Scale bar, 50 μm. (I) Oil Red O staining of liver tissue sections. Scale bar, 20 μm. (J) Serum levels of ALT and AST. (K) Relative mRNA expression of cytokines in the liver. (L) Representative images of Sirius Red staining and α-SMA immunostaining of liver tissue sections. Scale bar, 20 μm. The mRNA expression of genes was normalized to that of β-actin. Data are the mean ± SD; ns, not significant; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.