Extended Data Fig. 5: Attempts to degrade VHA-6 with the AID/AID2 system.
From: A lysosomal surveillance response to stress extends healthspan
a, The overall design for CRISPR/Cas9-mediated Degron::mNeonGreen knock-in with the endogenous tagging of a degron-mNeonGreen tag immediately after the last amino acid codon of vha-6 gene at chromosome II. b,c, The experimental flow (b) and genotyping result of the wild-type (+/+), heterozygous (+/KI) and homozygous (KI/KI) vha-6::Degron::mNeonGreen strains (c). d, The natural auxin indole-3-acetic acid (IAA) treatment at 1, 5 or 10 mM since egg stage barely reduced the expression level of Degron-mNeoGreen tagged VHA-6 in vha-6::Degron::mNeonGreen; eft-3p::TIR1::mRuby worms. vha-6 RNAi (20%) was used as a positive control. e, IAA at 1 mM remarkably decreased the Degron-mNeoGreen-DAF-16 expression in daf-16::Degron::mNeonGreen; eft-3p::TIR1::mRuby worms. f, 1-naphthaleneacetic acid (NAA) treatment at 1-10 mM since egg stage barely reduced the expression level of degron-mNeoGreen tagged VHA-6 in vha-6::Degron::mNeonGreen; eft-3p::TIR1::mRuby worms. g, The IAA derivative phenyl-indole-3-acetic acid (5-Ph-IAA) treatment at 5 or 10 μM since egg stage barely reduced the expression level of degron-mNeoGreen tagged VHA-6 in vha-6::Degron::mNeonGreen; eft-3p::TIR1(F79G)::mRuby worms. Scale bars, 0.2 mm.
