Extended Data Fig. 1: Kaem and Rhap induce mitophagy, abrogate disease pathologies, and improve healthspan and lifespan in cells, worm, or mouse models of AD.

a, b, electron microscopic images showing effects of Kaem and Rhap on mitochondrial morphology and mitophagy-like events in mt-Keima HeLa cells (a, 20 µM for 24 h) and mouse hippocampal brain tissues (b, 100 mg/kg/d via oral gavage from 12 months for 7 consecutive days; n = 3 mice per group with 4 random hippocampal neuronal images per mouse), treated with Kaem and Rhap. Blue arrows indicate mitophagosome-like events, while red arrows point to damaged mitochondria. Quantitative data of a and b are shown in Fig. 3m and Fig. 3n, respectively. c–f, Effects of Kaem and Rhap on the expression of designated proteins involved in Aβ production. N2a and N2S (N2a stably transfected with human Swedish mutant APP695) cells were used. One set of western blot data is shown (c), with semi-quantifications from three biological replicates (d–f). All quantitative data were shown in mean ± S.E.M. One-way ANOVA followed by Šidák’s multiple comparisons test (d-f) were used for data analysis. NS, no significance, *p < 0.05, **p < 0.01, ***p < 0.001. Original western blot gels for (c) are included in Source Data Fig. 2. g–i, Effects of Kaem and Rhap on lifespan of the N2 and hTau[P301L] (CK12) worms. Data shown are from one set of experiments from a total of two biological replicates (quantitative values in Supplementary Table 4). 90–120 worms were used for each group/biological repeat. All quantitative data were shown in mean ± S.E.M. Log-rank test was used for the statistics of lifespan data (g–i). NS, no significance, *p < 0.05, **p < 0.01, ***p < 0.001. j–l, Quantification of phosphorylated Tau sites (Thr181, Ser202/Thr205, Thr217) in hippocampal tissues from the 3xTg AD mice (n = 3 biological replicates). The western blotting gel data are shown in Fig. 6o. m–o, Effects of PINK1 knockdown on p-Tau inhibition by Kaem and Rhap in HEK 293 3G-EGFP-Tau P301L/mCherry cells. While one biological set of blot data are shown (m), quantifications (n, o) were from 3 biological replicates. All quantitative data were shown in mean ± S.E.M. Two-way ANOVA followed by Tukey’s multiple comparisons test (j, k. l) and one-way ANOVA followed by Šidák’s multiple comparisons test (n, o) were used for data analysis. NS, no significance, *p < 0.05, **p < 0.01, ***p < 0.001. Original western blot gels for (m) were included in Source Data Fig. 2.