A subset of C. elegans sHsps complements the growth deficits of yeast sequestrase mutants. (A) Fivefold serial dilutions of S. cerevisiae WT, hsp104∆ fes1∆ (∆∆), and ∆∆ btn2∆ cells expressing the indicated C. elegans sHsps were spotted on YPD agar plates and incubated at indicated temperatures for 3 d. Dashed lines indicate that spot tests from different plates are cropped and shown together for comparative analysis. (B) Yeast strains as in A were grown in YPD liquid media at 25°C. Log phase cultures were normalized to OD₆₀₀ = 0.2 and shifted to either 30 or 33°C. The OD₆₀₀ value for each strain at the time point when the hsp104∆ fes1∆ reference cell culture reached saturation was noted and plotted as a bar graph normalized to the OD₆₀₀ value corresponding to ∆∆btn2∆ cells without complementing C. elegans sHsp. A growth ratio of 1 denotes that the growth curves of ∆∆btn2∆ cells with and without C. elegans sHsp are equal. The dashed line corresponds to the set threshold (growth ratio 1.5). Error bars depict ±SD from n = 3 independent experiments. One-way ANOVA test (Dunnett) was used to assess the statistical significance of hits passing the set threshold (**, P < 0.01, ****, P < 0.0001). Data distribution was assumed to be normal, but this was not formally tested. (C) Venn diagram summarizing the results of C. elegans sHsp growth complementation tests of yeast sequestrase mutant cells (∆∆btn2 and ∆∆hsp42∆) both on solid agar and in liquid growth medium.