Fig. 3: Interaction between one-carbon metabolism and the RAS/MAPK pathway.

a, b Corpse and oocyte numbers for indicated genotypes grown at 15 °C and transferred to 25 °C 4 h before quantification, or grown at 20 °C and transferred to 25 °C 18 h. c DIC images of gonads of indicated genotypes grown at 20 °C and transferred to 25 °C 18 h before quantification. Numbers indicate mature oocytes. Scale bar: 10 µm. d Vulval induction index for indicated genotypes. a–d Animals were fed with E. coli OP50. e, f Corpse and oocyte numbers for indicated genotypes grown at 15 °C and transferred to 25 °C 4 h before quantification. g, h Corpse and oocyte numbers for indicated genotypes grown at 20 °C and transferred to 25 °C 5 h before quantification. e–h Animals were fed with E. coli OP50 (gray) or Comamonas DA1877 (blue). i dpERK intensity profiles in gonads of wild-type fed with E. coli OP50 (gray) or Comamonas DA1877 (blue), and let-60(gf ^ts) fed with E. coli OP50 (orange). x-axis represents normalized gonad length. Average intensities ± SD (shaded) are shown. Data from three independent biological replicates. j Western blot quantification of the somatic and germline isoform of dpERK for indicated genotypes. Bars represent mean ± SD normalized to the control (wild-type E. coli OP50) from three independent biological and technical replicates. See Supplementary Fig. 4 for the individual WB. k ERK-nKTR biosensor quantification. Dots represent individual animals from 2 independent biological replicates. Data show median ± min and max. l–m Normalized intensity of LIN-1::GFP or LIN-39::GFP at the Pn.px stage in 1° and 2° VPCs of let-60(gf) animals. Data were normalized to the control (E. coli OP50). Dots represent individual animals from two independent biological replicates. j–m Animals were fed with E. coli OP50 (gray) or Comamonas DA1877 (blue). Number of animals is in brackets. In a-h, j, l, m, boxplots show 25–75 percentiles and median; whiskers represent min and max values. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001 using one-way ANOVA followed by Dunnett’s multiple comparison tests in a and b, a Kruskal-Wallis test for non-parametric data in d, two-way ANOVA followed by a uncorrected Fisher’s LSD multiple comparison tests in e–h and j and unpaired, two-tailed t-tests in l and m. Source data are provided as a Source Data file.