Extended Data Fig. 2: Effect of vha-6 and vha-16 RNAi on gene expression, lifespan, stress responses and fertility.
From: A lysosomal surveillance response to stress extends healthspan
a, mCherry-VHA-6 expression of worms treated with control or 10%-60% vha-6 RNAi. b,c, GFP-VHA-16 expression (b) and survival (c) of worms treated with control or 10%-60% vha-16 RNAi (****P < 0.0001). d,e, GFP-VHA-16 expression (d) and survival (e) of worms treated with control or 0.1%-10% vha-16 RNAi (****P < 0.0001, *P = 0.0127 (ev VS vha-16 0.5%), P = 0.3713 (N.S., ev VS vha-16 0.1%)). f, RNAi of vha-6 attenuates the mitochondrial stress response activation induced by cco-1 RNAi in hsp-6p::gfp worms. RNAi targeting cco-1 occupies 40%, vha-6 RNAi occupies 20%. g, qRT-PCR analysis (n = 4 biologically independent samples) of worms as indicated in (f) (****P < 0.0001). h,i, RNAi of vha-6 attenuates the ER stress response induced by tunicamycin (5 μg/mL) (h) or hsp-3 RNAi (i) in hsp-4p::gfp worms. j, qRT-PCR analysis (n = 4 biologically independent samples) of worms as indicated in (i) (****P < 0.0001). k, RNAi of vha-6 attenuates the oxidative stress response induced by daf-2 RNAi in sod-3p::gfp worms. RNAi targeting daf-2 occupies 40%, vha-6 RNAi occupies 20%. l, qRT-PCR analysis (n = 4 biologically independent samples) of worms as indicated in (k) (****P < 0.0001). m, RNAi of vha-6 (20%) does not affect the heat shock response activation induced by heat shock (31 °C for 8 h). n, Different developmental stages of vha-6 RNAi treated worms all displayed much higher cpr-5p::gfp induction as compared to that in worms treated with control RNAi, suggesting that the LySR response and body size can be decoupled. o, The rate of egg-laying (n = 5 plates with 3 worms/plate for each condition) at different days of adulthood in worms treated with control, vha-6_1 or vha-6_2 RNAi. RNAi (100%) treatment started since the L4/young adult stage. The total egg output per worm was also calculated (****P < 0.0001, **P = 0.0018 (Day 4, ev VS vha-6_1), *P = 0.0277 (Day 4, ev VS vha-6_2)). Scale bars, 0.3 mm. Error bars denote SEM. Statistical analysis was performed log-rank test (c, e) and ANOVA followed by Tukey post-hoc test (g, j, l, o) (N.S., not significant). Statistical data for lifespan can be found in Supplementary Table 1.
