Fig. 3: MPP+ blocks autophagic flux as determined by an elevated calcium-dependent accumulation of insoluble p62 and Ub-positive spots. | Cell Death Discovery

Fig. 3: MPP+ blocks autophagic flux as determined by an elevated calcium-dependent accumulation of insoluble p62 and Ub-positive spots.

From: Buffering of cytosolic calcium plays a neuroprotective role by preserving the autophagy-lysosome pathway during MPP+-induced neuronal death

Fig. 3

MN9D cells were treated with 50 μM MPP+ in the presence or absence of 30 μM BAPTA-AM and/or 50 μM chloroquine (CQ) for 30 h. a Immunoblot analyses were performed using the anti-LC3 or anti-p62 antibody. b, c Quantification of LC3-II and p62 levels in each condition was performed after normalization to GAPDH loading control. Bars represent the mean ± SEM of three independent experiments. **p < 0.01; NS, not significant. After MPP+ treatment alone (d) or in combination with BAPTA-AM (e), cells were analyzed for the immunofluorescent localization of p62 (green) and LC3B (red) or ubiquitin (Ub; red). Cells were then examined using a confocal microscope. Merged views are provided in the right panel. The scale bar represents (d) 5 μm and (e) 10 μm. f A representative immunoblot analyses of p62, ubiquitin, and LC3 in Triton X-100 (TX)-soluble or TX-insoluble fractions

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