Fig. 3: Retromer stabilization rescues neuronal mtd/OXR1 deficiency in flies and human fibroblasts.
From: OXR1 maintains the retromer to delay brain aging under dietary restriction
a Retromer overexpression rescues developmental lethality induced by constitutively active neuronal mtdRNAi. n = at least 6 independent breeding chambers with 15 females to 5 males per chamber. Error bars represent mean value across replicates +/- SD. b Overexpression of vps26 rescues lifespan defects in flies with constitutively active neuronal mtdRNAi to the levels of flies under AL conditions. p value determined by log-rank test. c, Overexpression of vps35 rescues lifespan defects in flies with constitutively active neuronal mtdRNAi and restores DR-mediated lifespan extension. p value determined by log-rank test. d, R55 supplementation rescues developmental lethality induced by constitutively active neuronal mtdRNAi. n = at least 5 independent breeding chambers with 15 females to 5 males per chamber. Error bars represent mean value across replicates +/- SD. e 6 μM R55 supplementation throughout development and adulthood rescues lifespan defects in flies with constitutively active neuronal mtdRNAi and restores DR-mediated lifespan extension. p value determined by log-rank test. f 6 μM R55 supplementation throughout adulthood only extends lifespan. p value determined by log-rank test. g Human fibroblasts with a loss-of-function mutation in OXR1 (c.132delA) have reduced VPS35 levels, which is rescued by 10 µM R55 supplementation. Blue = DAPI, green = OXR1, red = VPS35. Quantification on right. n = minimum 12 cells across minimum 4 independent experiments. Error bars represent mean value across replicates +/- SD. h Immunocytochemistry in human fibroblasts with loss-of-function OXR1 mutation show enlarged endosomes (EEA1, red in top row), increased number of endosomes, and inability to traffic IGF2R to Golgi (GM130, red in bottom row). Quantification on right. n = minimum 38 cells across minimum 3 independent experiments. Error bars represent mean value across replicates +/- SD. For all figures, *p < 0.05, **p < 0.005, ***p < 0.0005. Except where noted, all p values were calculated by two-sided t-test.
