Fig. 3: Kaem and Rhap induce mitophagy in cells, C. elegans neurons and mouse brain.
a, Effects of Kaem and Rhap on the protein levels of mitofusin2 (MFN2), YFP-Parkin and Tim23 in HeLa cells stably overexpressing YFP-Parkin. b–d, Semi-quantification of a (n = 3 biological replicates). e, Effects of Kaem and Rhap on the protein levels of MFN2, YFP-Parkin and Tim23 in HeLa cells stably overexpressing Mito-GFP and mCherry-Parkin. f–h, Semi-quantification of e (n = 3 biological replicates). For a and e, CCCP was used as positive control. i, Images showing co-localization of mitochondria (Mito-GFP) and lysosomes (LAMP1 antibody) under Kaem and Rhap (20 uM, 24 h) administration in GFP-mito-mCherry-Parkin HeLa cells. White arrows indicate mitophagy events. j, Quantification of i with data from 3 biological repeats with around 5–7 images per biological repeat. k, Transgenic nematodes were treated with Kaem and Rhap (both with 0.01, 0.05, 0.1 and 0.2 mM), with mitophagy events calculated by the co-localization between the autophagic marker DsRed::LGG-1 and the mitophagy receptor DCT-1::GFP in neurons. n = 18–20 neurons from 2 biological repeats. While the left panel shows one representative set of images, quantitative data are shown in the right panel. l, Effects of Kaem and Rhap on the induction of neuronal mitophagy in worms with mt-Rosella reporter. Data were pooled from 2 biological replicates (total n = 20–35 nematodes per group), with the results shown as mean ± s.e.m. m,n, Data of quantified electron microscopic images showing effects of Kaem and Rhap on mitochondrial morphology and mitophagy-like events in mt-Keima HeLa cells (m) (20 µM for 24 h) and mouse hippocampal brain tissues (n) (100 mg kg−1 d−1 via oral gavage from 12 months for 7 consecutive days; n = 3 mice per group, with 4 random hippocampal neuronal images per mouse). Representative images are shown in Extended Data Fig. 1a,b. All quantitative data are shown as mean ± s.e.m. One-way ANOVA followed by Šidák’s multiple comparisons test; **P 0.01, ***P 0.001. Original unprocessed western blot gel data are in Source Data Fig. 4.
