Fig. 1: HSP-3 and HSP-4 show distinct spatial-temporal expression patterns and responses to ER-stress.
Representative western blot (a) and quantification (b) of PHX4377 (HSP-3::wrmScarlet) or PHX4415 (HSP-4::wrmScarlet) collected 24’, 48’, and 72’ after synchronization onto pos-1 siRNA. Membranes were probed with an anti-RFP (Chromotek, Cat #6g6) and anti-alpha tubulin (DSHB, Cat #12G10) antibody. n = 3 biological replicates. c Representative confocal images of PHX4377 and PHX4415 grown on pos-1 siRNA 24’, 48’, and 72’ after synchronization. Quantification (d) and representative confocal images (e) of PHX4377 grown on indicated siRNA and treated with 10 μg/mL of tunicamycin (Tm) or equivalent volume of M9 for 24 h. Quantification (f) and example confocal images (g) of PHX4415 grown on indicated siRNA and treated with 10 μg/mL of Tm or equivalent volume of M9 for 24 h. h Development assays of WT animals grown on indicated siRNA treated with M9 or 2.5 μg/mL of tunicamycin (Tm) as eggs. i Chronic Tm stress survival assay of WT worms grown on indicated siRNA transferred to plates treated with 20 μg/mL of Tm or equivalent volume of M9 as day 1 adults. M9 treatment: n = 110 (pos-1), n = 92 (hsp-3), n = 105 (hsp-4); Tm treatment: n = 133 (pos-1), n = 130 (hsp-3), n = 143 (hsp-4). For (c, e, g) images were taken at the midline and scalebars represent 100 μm. For (b, d, f, h) biological replicates (n = 3) are distinguished by color; triangles are average of technical replicates (circles). Source data are provided in the Source Data file. b, d–f, h Two-way ANOVA followed by Tukey’s multiple comparisons test. b pprotein x time = 0.0002; pprotein < 0.0001; ptime < 0.0001. d ptreatment x siRNA = 0.1; ptreatment < 0.0001; psiRNA < 0.0001. f ptreatment x siRNA < 0.0001; ptreatment < 0.0001; psiRNA < 0.0001. h ptreatment x siRNA < 0.0001; ptreatment < 0.0001; psiRNA < 0.0001. i Log-rank Mantel-Cox test. p < 0.05 is considered statistically significant. Error bars: s.d.
