Fig. 3: efk-1 functions independently of altering global translation rate.

A Fluorescence micrographs of irg-1p::GFP reporter worms (L4 stage) after RNAi against various translation machinery components or treatment with 2 mg/ml cycloheximide for 3 h. For RNAi experiments, empty vector RNAi-expressing HT115 E. coli (EV) is used as control. Data shown are representative of three independent experiments. Scale bar, 200 µm. B The graph shows L1 starvation survival of WT and efk-1 mutants with or without supplementation of 10 µM cycloheximide (CHX). N = 4, error bars represent mean ± SD; ****p < 0.0001 percent L4 vs. WT animals (AUC compared using one-way ANOVA with Tukey’s multiple comparisons test). C, D WB and quantification of puromycin incorporation in A549 lung cancer cells. Cells were treated with 1 µg/ml puromycin for 10 minutes. 1 h of 2 µg/ml CHX is used as control. N = 3, error bars represent mean ± SD; ***p = 0.0004, ****p < 0.0001 (two-way ANOVA with uncorrected Fisher’s LSD); for full membrane see Fig. S2L. E–H WB and quantification of puromycin incorporation in efk-1 mutants and WT both in E, F 8-h L4 starvation, and G, H 6-day L1 starvation. Worms were treated with 0.5 mg/ml puromycin for 3 h. 6 h of 2 mg/ml CHX is used as control. N = 3, error bars represent mean ± SD; for F, *p = 0.0232 for WT, *p = 0.0172 for efk-1 -CHX vs. +CHX; for H, *p = 0.0137 for WT, *p = 0.0441 for efk-1 -CHX vs. +CHX (two-way ANOVA with uncorrected Fisher’s LSD); for full membrane see Fig. S2M, N. WT, wild type; ns, not significant; AUC, area under the curve. Source data are provided as a Source Data file.