Fig. 1

HK attenuates lipid accumulation in lipotoxic hepatocytes through promoting autophagy. A Intracellular lipid content visualized with nile red (red) staining. Nuclei were stained with DAPI (blue). Scale bar = 10 μm. Resveratrol (10 μM) was used as a positive control. B The lipid content determined by Nile red staining. C The cellular TG content. D Atg5, Beclin1, LC3 and p62 protein levels in P/O-stimulated AML12 hepatocytes treated without or with various concentrations of HK or 10 µM resveratrol for 24 h. GAPDH was used as a loading control. E AML12 cells were infected with the Ad-mCheryy-GFP-LC3, and treated with or without HK. The mRFP-LC3 and GFP-LC3 puncta were visualized on a confocal microscope. Scale bar = 10 μm. F The numbers of autophagosomes (yellow puncta) and autolysosomes (red puncta) were quantitated. G In P/O-stimulated AML12 hepatocytes, HK treatment induced more LC3-positive puncta (green) on LDs that were visualized with Nile red staining (red). Scale bar = 25 μm. Rapamycin was used as a positive control. Data represented means ± SD. n = 6. *p < 0.05, **p < 0.01 and ***p < 0.001, HK or resveratrol vs. P/O treatment. ##p < 0.01, ###p < 0.001, ctrl vs. P/O treatment. One-way ANOVA was used to calculate the p-values