Fig. 3

mtROS disturbed lipid metabolism by impaired autophagy flux, which decreased PPARα/CPT1a and lipophagy. A, B Lipophagy was shown as the co-localization of LC3 (red) (A) or LTR (red) (B) with LDs indicated by Bodipy 493/503 (green). C Protein level of p62, LC3 I and LC3 II. D Cells transfected with mRFP-GFP-LC3 adenovirus were treated with CES and MitoQ for 24 h. Then, pictures were captured. Yellow dots were autophagosomes and red were autolysosomes. E Protein level of collagen I, α-SMA, PPARα and CPT1a in each group. F Lipid content was assessed by Nile Red staining. *p < 0.05 versus control, ^#p < 0.05 versus CSE, ^&p < 0.05 versus CSE + MitoQ (n = 3). G Protein level of collagen I and α-SMA of cells treated with ETO or OA alone for 24 h. H Nile Red staining was performed to examine lipid content of cells stimulated with ETO or OA for 3 h. I Cells pretreated with or without BA for 1 h were treated with ETO or OA for 24 h. Then lipid content was evaluated by Nile Red staining. J Autophagosomes (yellow) and autolysosomes (red) were observed. K Cells pretreated with or without BA for 1 h were stimulated with ETO or OA for 3 h. Then protein level of LC3 I and LC3 II were detected. *p < 0.05 versus control, ^#p < 0.05 versus ETO, ^&p < 0.05 versus OA (n = 3). BA bafilomycin, LTR lysotracker red