Fig. 2: NAC pretreatment alleviated the AOPP-induced inhibitory effects on autophagy and lysosomal biogenesis in granulosa cells. | Cell Death & Disease

Fig. 2: NAC pretreatment alleviated the AOPP-induced inhibitory effects on autophagy and lysosomal biogenesis in granulosa cells.

From: Advanced oxidation protein products attenuate the autophagy-lysosome pathway in ovarian granulosa cells by modulating the ROS-dependent mTOR-TFEB pathway

Fig. 2

A A DCF fluorescence assay showing the intracellular ROS production in negative control, KGN/COV434 cells treated with 200 μg/mL AOPPs for 48 h (the AOPP group), cells treated with 2 mM NAC for 49 h (the NAC group), and cells pretreated with 2 mM NAC for 1 h and then treated with AOPPs for 48 h (the NAC + AOPP group). B Representative blots and quantification showing p62 and LC3B II expression levels in the control group, AOPP group, NAC group, and NAC + AOPP group. C Representative images of RFP-GFP-LC3 under a confocal laser scanning microscope in the control group, AOPP group, NAC group, and NAC + AOPP group. The GFP + RFP+ (yellow) puncta are autophagosomes. The numbers of autophagosomes were quantified for 10 cells. High-magnification images of the indicated boxed areas are shown in the bottom panels. D Lysosomal biogenesis assessed by Lyso-Tracker Red in the control group, AOPP group, NAC group, and NAC + AOPP group. E LAMP1 expression as detected by immunofluorescence in the control group, AOPP group, NAC group, and NAC + AOPP group. All experiments were repeated for three times. The results are expressed as the mean ± SD from experiments performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001. Statistical significance was determined by one-way ANOVA and Tukey tests for post hoc comparisons.

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