Fig. 7: Hsf1 promotes autophagy and alleviates hepatic steatosis caused by Usp39 deletion.

a, b AML12 cells transfected with indicated vectors. Cells were stained with BODIPY and imaged using confocal microscopy. Lipid droplet numbers were quantified of 30 BODIPY-stained cells in each group (n = 3 independent experiments) by Image J. Scale bar, 10 µm. c Co-localization between Plin2 (green) and Lamp2 (red) was analyzed by immunofluorescence staining in AML12 cells transfected with indicated vectors, co-localization signal was quantified by Image J. Scale bars, 10 µm. d Immunoblotting analysis of Usp39, Hsf1 and autophagy-related genes in AML12 cell transfected with indicated vectors in complete or serum deprived medium (n = 3, 4 per group, from three independent experiments). e Oil Red O staining of liver sections of Usp39-floxed mice injected with AAV-empty, AAV-TBG-Cre, or Ad-Hsf1. The mice were sacrificed after fasted for 16 h. Scale bars, 50 µm. f Immunofluorescence experiments showed co-localization of Plin2 (green) and Lamp2 (red). Scale bar, 10 µm. g Immunoblotting analysis of protein levels of indicated genes in the livers from Usp39-floxed mice injected with AAV- empty, AAV-TBG-Cre, or Ad-Hsf1. The mice were sacrificed after fasted for 16 h (n = 3, 4, 6 per group, from three independent experiments). Immunoblotting band intensities were quantified by Image J, which were shown in Fig. S10d, j. Co-localization was quantified by Image J. Images are representative of at least three independent experiments. P-value was obtained by unpaired two-sided Student’s t-test and the results are presented as the mean ± S.D. NS, stands for non-significant. AD, Adenovirus. AAV, Adeno-associated virus. TBG, thyroxine-binding globulin. MW, molecular weight. Source data are provided as a Source data file.