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. 2023 Jul 25;12:e83884. doi: 10.7554/eLife.83884

Figure 1. Knockdown of let-767 specifically suppresses the UPRER.

(A) Schematic for screening method used to identify UPRER modulators from candidate genes. Animals expressing hsp-4p::GFP were grown from L1 on candidate RNAi mixed in a 1:1 Ratio with ER stress inducing sec-11 RNAi. Animals were then screened at day 1 of adulthood and scored for changes in fluorescence compared to the sec-11/Empty Vector (EV) control. (B) Fluorescent micrographs of day 1 adult transgenic animals expressing hsp-4p::GFP grown from L1 on EV, sec-11, or tag-335 RNAi combined in a 1:1 ratio with either EV or let-767 RNAi to assay effects on UPRER induction. (C) Quantification of (B) normalized to size using a BioSorter. Lines represent mean and standard deviation. n=500. Mann-Whitney test p-value ****<0.0001. Representative data shown is one of three biological replicates. (D) Fluorescent micrographs of day 1 adult transgenic animals expressing hsp-16.2p::GFP grown from L1 on EV or let-767 RNAi with or without 2 hr 34 °C heat-shock treatment to assay heat shock response. Animals imaged 2 hr after recovery at 20 °C. (E) Quantification of (D) normalized to size using a BioSorter. Lines represent mean and standard deviation. n=400. Mann-Whitney test n.s.=not significant. Representative data shown is 1 of 3 biological replicates. (F) Fluorescent micrographs of day 1 adult transgenic animals expressing hsp-6p::GFP, grown from L1 on EV or cco-1 RNAi combined in a 1:1 ratio with either EV or let-767 RNAi to assay effects on UPRmt induction. (G) Quantification of (F) normalized to size using a BioSorter. Lines represent mean and standard deviation. n=431. Mann-Whitney test p-value ****<0.0001. Representative data shown is one of three biological replicates.

Figure 1.

Figure 1—figure supplement 1. Knockdown of let-767 suppresses the UPRER more severely than other lipid related genes.

Figure 1—figure supplement 1.

(A) Fluorescent micrographs of transgenic animals expressing hsp-4p::GFP grown from L1 on Empty Vector (EV) or sec-11 RNAi combined in a 1:1 ratio with either atp-1, atp-2, rab-1, fib-1, tba-2, let-767, hsp-90, hsp-1, ucr-1, vha-13, or vha-15 RNAi and imaged at day 1 of adulthood to assay effects on UPRER induction. (B) Quantitative RT-PCR transcript levels of xbp-1s, total xbp-1, and let-767 from day 1 adult N2 animals grown from L1 on EV or sec-11 RNAi combined in a 1:1 ratio with either EV or let-767 RNAi. Fold-change compared to EV treated N2 animals. Error bars indicate ± standard deviation across three biological replicates, each averaged from two technical replicates. (C) Fluorescent micrographs of day 1 adult transgenic animals expressing hsp-4p::GFP grown from L1 on EV, stdh-1, stdh-2, stdh-3, or stdh-4 RNAi combined in a 1:1 ratio with either EV or sec-11 RNAi to assay effects on UPRER induction. (D) Quantification of (C) normalized to size using a BioSorter. Lines represent mean and standard deviation. n=245. Mann-Whitney test p-value **<0.05, ****<0.0001. Representative data shown is one of three replicates. (E) Fluorescent micrographs of transgenic animals expressing hsp-4p::GFP grown from L1 on EV, pod-2, acs-1, hpo-8, or elo-5 RNAi combined in a 1:1 ratio with either EV or sec-11 RNAi to assay effects on UPRER induction. (F) Quantification of (E) normalized to size using a BioSorter. Lines represent mean and standard deviation. n=236. Mann-Whitney test p-value ****<0.0001. Representative data shown is one of three biological replicates.