Fig. 7: Blockage of autophagy rescues age-related apoptosis of LECs through preventing p62 degradation.

A Representative confocal images of primary cultured human LECs from healthy donors of different ages stimulated with 50 μM H₂O₂ for indicated times. LECs are stained with LC3 antibody (red), p62 antibody(cyan), TUNEL labeling (green), and DAPI (blue). Scale bar, 30 μm. B Fluorescent intensity of p62 in primary cultured human LECs from healthy donors treated as in A (mean ± SD; n = numbers of healthy donors analyzed, ****p < 0.0001, Kruskal-Wallis test followed by Bonferroni post hoc test). C Correlation between p62 fluorescence intensity and the number of LC3 puncta in LECs from healthy donors of different age groups treated as in A (100 cells were analyzed; each symbol represents one LEC). p = 0.000338 by Spearman’s rank correlation test. D Representative confocal images of primary cultured human LECs from elder healthy donors pretreated with or without 0.5 mM 3-MA for 3 h, followed by 50 μM H₂O₂ exposure for indicated times. LECs are stained with LC3 antibody (red), p62 antibody(cyan), TUNEL labeling (green), and DAPI (blue). Scale bar, 30 μm. E Fluorescent intensity of p62 in primary cultured human LECs from elder healthy donors treated as in D. (mean ± SD; n = numbers of healthy donors analyzed, ****p < 0.0001, Mann-Whitney U Test). F Grouped stacked bars showing the percentages of LC3^-, LC3⁺, LC3^{+ +}, and LC3⁺⁺⁺ in primary cultured human LECs from elder healthy donors treated as in D (mean ± SD; n = numbers of healthy donors analyzed; *p < 0.05, ***p < 0.001, Mann-Whitney U Test). G Percentages of TUNEL-positive primary cultured human LECs from healthy donors treated as in D (mean ± SD; n = numbers of healthy donors analyzed; ****p < 0.0001, two-way ANOVA followed by Bonferroni post hoc test).