Fig. 8: Targeted screen to identify genes involved in the co-regulation of lipid droplet and peroxisome number.
a, Number of intestinal lipid droplets and peroxisomes, measured by fluorescence, in dhs-3p::dhs-3::GFP; vha-6p::mRFP–SKL worms during aging and following ash-2 depletion. Data are the mean ± s.e.m. of n = 21–37 worms. Each dot represents the mean organelle number of all worms imaged for this condition. b, Targeted screen design. Created with BioRender.com c, Genes involved in the co-regulation of organelle number. Number of intestinal lipid droplets and peroxisomes, measured by fluorescence, in dhs-3p::dhs-3::GFP; vha-6p::mRFP–SKL worms following treatment with 62 different RNAis. RNAi of the indicated genes resulted in an increase (red) or decrease (teal) of both organelles; n ≥ 18 worms per condition. d, Number of intestinal lipid droplets and peroxisomes, quantified by fluorescence, in dhs-3p::dhs-3::GFP; vha-6p::mRFP–SKL worms following sbp-1 and nhr-49 transcription factor depletion; n = 24–30 worms. e, Number of intestinal lipid droplets and peroxisomes, quantified as in d, following ash-2, fat-2 or hosl-1 depletion; n = 20–42 worms. f, Genes involved in uncoupling lipid droplet and peroxisome number. RNAi of the indicated genes resulted in decreased numbers of lipid droplets and increased peroxisomes (yellow) or increased numbers of lipid droplets and decreased peroxisomes (blue). g, Number of lipid droplets and peroxisomes, measured as in d, following vps13d depletion; n = 33 or 34 worms. h, Number of lipid droplets and peroxisomes, measured as in d, following rab-7 depletion; n = 29–41 worms. d,e,g,h, Data are the mean ± s.d. Each dot represents the organelle number of an individual worm normalized to control worms. P values were determined using a two-tailed Mann–Whitney test. i, Genes important for lifespan. Perturbations that affect the co-regulation of the number of lipid droplets and peroxisomes, color-coded according to their effect on lifespan and MUFA-induced longevity. c,f,i, Each dot represents the mean organelle number, normalized to control, of all worms imaged for this condition. A two-tailed Pearson’s R2 test was used to analyze correlation; dotted line, nonlinear fit. j, vps13d is not necessary for longevity following ash-2 depletion; n ≥ 71 worms for each condition. k, rab-7 is necessary for longevity following ash-2 depletion; n ≥ 93 worms for each condition. j,k, Percentages of the median lifespan extension and P values, determined using a log-rank Mantel–Cox test, are indicated. a,d,e,g,h,j,k, Data are representative of three (d,g,h,j,k) or two (a,e) independent experiments. Source data are provided.
