Figure 7

Effect of ZK treatment on LC3-I and LC3-II protein expression in polarized ARPE-19 cells pretreated with H₂O₂. RLUs of (a) LC3-I and (b) LC3-II normalized to the amount of total loaded protein in polarized ARPE-19 cells prior to H₂O₂ exposure and polarized ARPE-19 cells challenged with 300 µM H₂O₂ for 3 h or 24 h followed by a resting period (control) or treatment with 10–100 nM ZK for 3 h. (c) Representative western blot analysis showing total protein loading and LC3-I and LC3-II in polarized ARPE-19 cells prior H₂O₂ exposure and polarized ARPE-19 cells challenged with H₂O₂ for 3 h or 24 h followed by ZK treatment for 3 h. ARPE-19 cells were cotreated with 10 µg/ml E64d and pepstatin A (lysosomal degradation inhibitors) to prevent LC3-II degradation upon H₂O₂ challenge. The data are presented in box and whisker plot format (min to max); n = 10. Significance was calculated by repeated measures two-way ANOVA (main factors: H₂O₂ exposure (matched) and ZK treatment (matched); interaction H₂O₂ exposure × ZK) followed by Bonferroni’s multiple comparison test. **p < 0.01, ***p < 0.001.