Fig. 4

Effects of PGF2α and PGE2 on autophagy and lipid accumulation in zebrafish liver cells. Cells were incubated with or without PGF2α (10 μM) and PGE2 (10 μM) for 24 h. (a) Lipid droplets (LDs) were stained with BODIPY (green), nuclei were stained with DAPI (blue), lysosomes were stained with LysoTracker (red), and LC3A/B was stained with a specific immunofluorescent antibody (red). (b–e) Triglyceride (TG) content (n = 4), relative fluorescence of lysosome puncta (n = 3) and LC3A/B (n = 3), and Mander’s overlap coefficient of lysosome/BODIPY (n = 3) in each image were quantified using ImageJ software (n = 4). (f) Relative transcript expression of autophagy-related genes (n = 3). (g) Cells were pre-incubated with autophagy inhibitor CQ, then treated with PGF2α for 24 h; the LDs and nuclei were stained with BODIPY and DAPI, respectively. ATG12, autophagy-related 12; EF1α, elongation factor 1 α; LAMP2, lysosomal-associated membrane protein 2; LC3b, microtubule-associated protein 1 light chain 3b. Statistical significance is denoted with asterisks as follows: * p < 0.05; ** p < 0.01.