Analysis of ASNA-1 million mutation point mutants.

(A) Multiple sequence alignment of ASNA-1/GET3/TRC40 comparing particular domains of the protein in 4 different species. The essential domains are highly conserved. Amino acids mentioned throughout the paper are marked in blue. (B) Confocal imaging merge of 1-day old adults co-expressing mCherry::SP12 with either ASNA-1::GFP or ASNA-1A63V::GFP. (C) Expression from the hsp-4p::GFP reporter (zcIs4) imaged by fluorescence microscopy in the 1-day old adult wild-type and asna-1(A63V) animals. hsp-4p::GFP expression quantification in the wild-type (n = 5) and asna-1(A63V) animals (n = 5). Statistical significance was determined by the independent two-sample t-test. Bars represent mean ± SD. (D) Relative mRNA analysis of ER stress reporter hsp-4 in 1-day old adult asna-1(A63V) animals. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Relative mRNA analysis of (E) the mitochondrial stress reporters (hsp-6 and hsp-60) and (F) oxidative stress reporters (gst-4, gst-30, and gst-38) in 1-day old adult wild-type and asna-1(A63V) animals. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. (TIFF)