Fig. 2: ATF-4 mediates lifespan extension from translation inhibition.

a Adult-specific knockdown of ifg-1 extended the lifespan of WT animals but not atf-4(tm4397) mutants. b Adult-specific treatment with 25 μM cycloheximide increased lifespan dependent upon atf-4. c Representative western blots and quantification (d) showing that treatment with 35 μg/ml tunicamycin for 4 h dramatically increased eIF2α phosphorylation levels in L4 stage animals, while treatment with 7.2 mM cycloheximide for 1 h did not. Mean ± SEM. n = 3 biological replicates. One-way ANOVA with post hoc Tukey. e Representative western blots and quantification (f) showing the effects of adult-specific knockdown of eif-1.A, ifg-1, ragc-1, or rict-1 on eIF2α phosphorylation levels. Mean ± SEM. n = 3 biological replicates. One-way ANOVA with Dunnett’s post-test compared to EV. g Representative western blots of puromycin incorporation assay and quantification (h) showing that adult-specific knockdown of ifg-1, ragc-1, or rict-1 decreased translation. Mean ± SEM. n = 3 biological replicates. One-way ANOVA with Bonferroni post-test. i Quantification of GFP fluorescence showing that adult-specific ifg-1 knockdown increases expression of Patf-4(uORF)::GFP. j Adult-specific knockdown of ifg-1 comparably extended the lifespan of WT animals and eif-2α/y37e3.10(qd338) phosphorylation-defective mutants. For statistics and additional trials in (a), (b), and (j), see Supplementary Data 1. For western blots, source data are provided in Source data File.