Fig. 3: ELT-2 regulates LySR activation and LySR-associated lifespan extension.
From: A lysosomal surveillance response to stress extends healthspan
a, The most enriched binding motif in the promoters of vha-6 RNAi only 760 genes. The P value was derived from HOMER (a one-sided hypergeometric test). b, The genomic distribution of the motif hits at promoters of vha-6 RNAi only and all other genes. c, RNAi of elt-2 attenuated GFP expression of cpr-5p::gfp worms upon vha-6 RNAi. Scale bar, 0.3 mm. d,e, The western blots (d) and qRT–PCR analysis (e) (n = 4 biologically independent samples) of cpr-5p::gfp worms treated with control, vha-6 and/or elt-2 RNAi (****P < 0.0001; for cpr-5, ***P = 0.0006 (elt-2 versus elt-2 + vha-6); for ctsa-1, P = 0.1563 (not significant (n.s.), control (ev) versus elt-2), *P = 0.0136 (elt-2 versus elt-2 + vha-6); for asp-10, P = 0.2298 (n.s., ev versus elt-2), P = 0.0798 (n.s., elt-2 versus elt-2 + vha-6); for elt-2, ***P = 0.0002 (ev versus vha-6), ***P = 0.0003 (ev versus elt-2), P = 0.1278 (n.s., elt-2 versus elt-2 + vha-6)). f, A qRT–PCR analysis (n = 4 biologically independent samples) of worms treated with indicated RNAi (****P < 0.0001; for cpr-5, P > 0.9999 (n.s., elt-2 versus elt-2 + vha-8/vha-14/vha-15), P = 0.9999 (n.s., elt-2 versus elt-2 + vha-20); for cpr-8, P > 0.9999 (n.s., elt-2 versus elt-2 + vha-8), P = 0.9971 (n.s., elt-2 versus elt-2 + vha-14), P = 0.9977 (n.s., elt-2 versus elt-2 + vha-15), P = 0.9706 (n.s., elt-2 versus elt-2 + vha-20)). g, A Venn diagram of DEGs with indicated conditions. h,i, A functional clustering of the 318 (h) and 1,229 (i) DEGs in g. The P value was derived from DAVID (a one-sided Fisher’s exact test). j,k, Western blots (j) and ChIP–qPCR (k) (n = 4 biologically independent samples) of elt-2p::elt-2::gfp-flag worms treated with indicated RNAi (****P < 0.0001). l, The survival of worms treated with indicated RNAi (****P < 0.0001, P = 0.0541 (n.s., elt-2 versus elt-2 + vha-6)). The error bars denote the standard error of the mean. The statistical analysis was performed by ANOVA followed by Tukey’s post hoc test in e, f and k or a log-rank test in l. The statistical data for lifespan can be found in Supplementary Table 1.
