Figure EV4. Lifespan effect of RET inhibition by CPT is mediated by Sirtuin, Foxo, and autophagy pathways.

• A, B
  Immunoblots showing the effect of aging on dFoxo acetylation (A), and effect of CPT treatment on dFoxo acetylation (B). dFoxo was immunoprecipitated from whole fly extracts and detected with anti‐acetyl‐K antibody.
• C
  RT–PCR analysis of levels of the indicated transcripts in aged flies relative to young flies.
• D
  Images and data quantification showing p62 staining in the brain of old flies and old flies treated with CPT (n = 5 per group).
• E–I
  Survival curve showing the effect of NMN on lifespan in control (Mhc > wt) flies (**P < 0.01, logrank) (E), and NDUFS3‐RNAi (F), Foxo‐RNAi (**P < 0.01, logrank) (G), Sirt‐2‐RNAi (H), and ATG1‐RNAi (**P < 0.01, logrank) (I) flies (n = 3 groups, 20 flies per group).

Data information: Data are representative of at least three repeats. Data are shown as mean ± SEM in (C, D). Survival curves were analyzed with logrank test along with Wang–Allison test for maximum lifespan. Dashed lines mark time points of 50% survival. Asterisks indicate statistical significance (***P < 0.001, **P < 0.01, *P < 0.05) in single‐factor ANOVA with Scheffe's analysis as a post hoc test.