Fig. 2: Mutations in ppp-1 do not attenuate mRNA translation. | Nature Communications

Fig. 2: Mutations in ppp-1 do not attenuate mRNA translation.

From: Mutagenesis screen uncovers lifespan extension through integrated stress response inhibition without reduced mRNA translation

Fig. 2

a Puromycin incorporation followed by Western blot analysis using antibodies detecting puromycin and α-tubulin in day 1 WT animals, iftb-1(wrm53) mutants, and control rsks-1(sv31) mutants. Images of the same membrane are shown with short and long exposures (error bars represent means + SEM, one-way ANOVA Dunnett’s post hoc test with **p = 0.0023 iftb-1(wrm53) vs. WT and ***p < 0.001 rsks-1(sv31) vs. WT; n = 3 independent experiments). b Puromycin incorporation assay in day 1 WT animals, ppp-1 mutants, and rsks-1(sv31) controls as described in (a). Images of the same membrane are shown with short and long exposures (error bars represent means + SEM, one-way ANOVA Dunnett’s post hoc test with ***p < 0.001 vs. WT; n = 6 independent experiments). c Quantification of methionine 35S labeling of day 1 WT worms, ppp-1 mutants, and control rsks-1(sv31) mutants (error bars represent means + SEM, one-way ANOVA Dunnett’s post hoc test with ***p < 0.001 vs. WT; the number of independent experiments (n) indicated in the figure). d, e Polysome profiling, and quantification of day 1 WT and ppp-1 animals. Quantification represents the relative abundance of ribosomal subunits (40S, 60S), monosomes (mono), and polysomes (poly; error bars represent means + SD, two-way ANOVA Dunnett’s post hoc test; n = 4 independent experiments). Source data are provided as a Source Data file.

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