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. 2022 Jan 10;8:1. doi: 10.1038/s41420-021-00796-3

Fig. 3. APP-CTFs accumulation triggers mitophagy dysfunction in AD-iNSCs.

Fig. 3

A Representative western blotting showing LC3 and p62 expression in WT- and AD-iNSCs treated or not with bafilomycin or rapamycin for 6 h. B Quantification of LC3-II, LC3-II/LC3-I, and the p62 ratio in WT- and AD-iNSCs with respect to control iNSCs under vehicle-treated conditions. C Representative images showing LC3 and LAMP1 in WT- and AD-iNSCs under basal conditions. D Representative western blotting showing APP, APP-CTFs, LC3, p62, Parkin, PINK1, HSP60, and β-actin expression in AD-iNSCs treated or not with DAPT for 24 h. E Quantification of autophagy-related markers, including LC3 and p62, in the presence of DAPT relative to vehicle treatment. F Quantification of APP-CTFs and mitophagy-related markers, including Parkin, PINK1, and HSP60, in the presence of 10 µM DAPT relative to vehicle treatment. G Representative z-stack images showing APP-CTF in green and HSP60 as a mitochondrial membrane marker in red in AD-iNSCs treated with 10 µM DAPT for 24 h. Scale bars, 20 µm. H Representative images showing LAMP1 (Lysosome marker) and MitoTracker (Mitochondria marker) in AD-iNSCs treated or not with DAPT for 24 h. Scale bars, 10 µm. I Quantification of colocalization area with LAMP1 and MitoTracker. Statistical analysis was performed by Student’s t-test. n.s not significant, *P < 0.05, **P < 0.01 and ***P < 0.001. The results are presented as the means ± SD.