Extended Data Fig. 9: Role of lysosomal proteases in LySR activation and vha-6 RNAi-induced beneficial effects.
From: A lysosomal surveillance response to stress extends healthspan
a-c, Knockdown validations with qRT-PCR analysis (n = 4 biologically independent samples) of worms treated with RNAi as indicated (****P < 0.0001). d-g, Worms expressing either unc-54p::Q35::YFP (polyQ model) (d and e) or unc-54p::Hsa-sod-1::YFP (ALS model) (f and g) were treated with control, cpr-5 and/or vha-6 RNAi, and analyzed at Day 5 of adulthood (n = 10 individual worms for each condition). RNAi targeting cpr-5 occupied 80%, vha-6 RNAi occupied 20%. Control RNAi was used to supply to a final 100% of RNAi for all conditions (****P < 0.0001; in (e), P = 0.9999 (N.S., ev VS cpr-5), **P = 0.0037 (vha-6 VS vha-6 + cpr-5); in (g), P = 0.9999 (N.S., ev VS cpr-5)). Scale bars, 0.3 mm. h, Transgenic worm strains expressing elt-2 promoter-driven mCherry and Degron-mNG-tagged ELT-2. Scale bars, 0.1 mm. i, The expression pattern of intestine-specific ges-1 promoter-driven DsRed-CPR-5 and GFP in ges-1p::cpr-5::DsRed;SL-2::GFP worms. The CPR-5-DsRed is localized not only in the intestine but also in the six coelomocytes (white arrows and in dashed circles). The gene expression of cpr-5 is indicated by polycistronic GFP, while the CPR-5 protein is visualized by its RFP fusion. Scale bar, 100 μm. j, The level of secreted CPR-5-RFP fusion is increased by vha-6 RNAi, but not by vha-16 or vha-19 RNAi (n = 15 individual worms for each condition) (****P < 0.0001, P = 0.8769 (N.S., ev VS vha-16), P = 0. 9843 (N.S., ev VS vha-19)). Error bars denote SEM. Statistical analysis was performed by ANOVA followed by Tukey post-hoc test (N.S., not significant).
