Fig. 6

Pivotal role of ASM in the regulation of HG-induced autophagy for the execution of ferroptosis. hFob1.19 cells were treated with HG (35 mM) in the presence or absence of ASM-shRNA, HCQ (10 µM) and NAC (5 mM). Following the treatment, A ASM expression was determined using Western blot analysis. B Ceramide levels were measured. C Western blot analysis of LC3 II was carried out. D Western blot analysis of indicated proteins were carried out. E Autophagic flux was determined by ptfLC3 fluorescent probe. Scale bar = 20 μm. F LPO by C11 BODIPY 581/591 fluorescent probe was assessed. Scale bar = 20 μm. G ROS generation was detected by DHE assay kits. Scale bar = 20 μm. H LPO was determined by MDA assay. I LPO was detected by the 4-HNE assay kits. All data are presented as the mean ± SD of three independent experiments.****P < 0.0001