Extended Data Fig. 4: Auxin-inducible degradation (AID) of ELT-2 mimics the effect of elt-2 RNAi in LySR and lifespan determination.
From: A lysosomal surveillance response to stress extends healthspan
a, The overall design for CRISPR/Cas9-mediated elt-2::Degron::mNeonGreen knock-in with the endogenous tagging of a Degron-mNeonGreen tag immediately after the last amino acid codon of the elt-2 gene at chromosome X. b, The genotyping results of the wild-type, homozygous and heterozygous elt-2::Degron::mNeonGreen knock-in strains. c, The natural auxin indole-3-acetic acid (IAA) treatment at 0.1-0.8 mM remarkably decreased the Degron-mNG-ELT-2 expression in the elt-2::Degron::mNeonGreen; eft-3p::TIR1::mRuby (ELT-2 AID) worms. Scale bars, 0.1 mm. d, qRT-PCR analysis (n = 4 biologically independent samples) of the ELT-2 AID worms treated with control (ctrl) or 0.1 mM IAA (****P < 0.0001; for cpr-5, P = 0.8422 (N.S., ev + IAA VS vha-6 + IAA); for cpr-8, P = 0.8689 (N.S., ev + IAA VS vha-6 + IAA); for ctsa-1, P = 0.6185 (N.S., ev + IAA VS vha-6 + IAA); for asp-10, P = 0.3410 (N.S., ev + IAA VS vha-6 + IAA); for elt-2, **P = 0.0012 (ev VS vha-6), **P = 0.0043 (ev VS ev + IAA), **P = 0.0033 (ev + IAA VS vha-6 + IAA)). e, Survival of ELT-2 AID worms treated with control or vha-6 (20%) RNAi and/or 0.1 mM IAA (****P < 0.0001, **P = 0.0022 (ev VS ev + IAA)). Error bars denote SEM. Statistical analysis was performed by ANOVA followed by Tukey post-hoc test (d) or log-rank test (e) (N.S., not significant). Statistical data for lifespan can be found in Supplementary Table 1.
