Fig. 3: Autophagy facilitates apoptosis of LECs independent of ROS.

A Representative ROS levels in WT and ATG7 KO HLE-B3 cells pretreated with 10 mM NAC for 1 h, followed by 200 μM H₂O₂ stimulation for 24 h assessed by flow cytometry using the ROS sensor DCFH-DA (left). Right panel, quantitative data from four independent experiments (mean ± SD; *p < 0.05, **p < 0.01, unpaired Student’s t test). MDA (B), SOD (C), and GSH (D) levels in WT, ATG7, or ATG3 KO HLE-B3 cells with or without 200 μM H₂O₂ exposure for 24 h measured by respective kits (MDA level determined by TBRAS assay and normalized to protein concentration). Quantitative data are presented as mean ± SD from four independent experiments (**p < 0.01, H₂O₂-treated vs control group; two-way ANOVA followed by Bonferroni post hoc test). E Viability of WT and ATG7 KO HLE-B3 cells pretreated with or without 10 mM NAC for 1 h, followed by 200 μM H₂O₂ stimulation for 24 h determined by CCK8 assay. Mean ± SD from four independent experiments; ****p < 0.0001 (unpaired Student’s t test). F MitoTracker labeling and FACS analysis of mitochondrial mass in WT and ATG7 KO HLE-B3 cells with or without 200 μM H₂O₂ exposure for 24 h. Right panel, quantitative data from four independent experiments (mean ± SD; **p < 0.01, ****p < 0.0001, unpaired Student’s t test).