Fig. 5: Overexpression of TFEB alleviated the AOPP-induced inhibitory effects on autophagy and lysosomal biogenesis in granulosa cells. | Cell Death & Disease

Fig. 5: Overexpression of TFEB 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. 5

A Immunofluorescence staining of TFEB in negative control cells, KGN/COV434 cells treated with 200 μg/mL AOPPs for 48 h (AOPP group), cells transfected with LV-NC (LV-NC group), cells transfected with LV-NC and treated with AOPPs (LV-NC + AOPP group), cells transfected with LV-TFEB (LV-TFEB group), and cells transfected with LV-TFEB and treated with AOPPs (LV-TFEB + AOPP group). B Representative blots and quantification showing p62 and LC3B II expression levels in the control group, AOPP group, LV-TFEB + AOPP group and LV-NC + AOPP group. C Representative images of RFP-GFP-LC3 under a confocal laser scanning microscope in KGN cells in the control group, AOPP group, LV-TFEB + AOPP group and LV-NC + 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 LysoTracker Red in the control group, AOPP group, LV-TFEB + AOPP group and LV-NC + AOPP group. E LAMP1 expression as detected by immunofluorescence in the control group, AOPP group, LV-TFEB + AOPP group and LV-NC + 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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