Fig. 4: Thiols shorten C. elegans lifespan by inhibiting SKN-1-dependent transcription.

a Regulation of representative SKN-1 dependent genes by LB, DB or DB + NAC. Graphs show RPKM of three independent experiments from RNA-seq. See Supplementary Data 1 for statistics. b, c Representative fluorescent image and quantification demonstrating that skn-1-dependent transcription is downregulated by thiols. L4 stage gst-4::GFP worms transferred to live or dead bacteria diet ±15 mM NAC and incubated for 5 days prior to imaging. n = 60–65 worms over three independent experiments. See also Supplementary Table 5. d NAC (5 mM) does not shorten the lifespan of skn-1 worms. Average percentage change ±SD of the lifespan relative to untreated control is indicated in red (n = 188(untreated), 177(5 mM NAC)). e, f Representative fluorescent image and quantification demonstrating ROS staining by DHE in live worms. WT animals were reared on LB, DB or DB + 15 mM NAC and stained with DHE at day 3 of adulthood. n = 91–94 worms over three independent experiments. See also Supplementary Table 6. In a, c, f the box plots indicate median (middle line), 25th, 75th percentile (box) and 5th and 95th percentile (whiskers) as well as maximum, minimum and mean (single points). In all graphs p values are: n.s. not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; two-tailed t-tests.