Fig. 2: MET-2 deficient in H3K9 HMT activity forms foci and promotes germline and somatic development.
a, Allele map of met-2 locus showing deletion allele met-2Δ(n4256) as black bar and point mutant met-2-CD(gw1660). Amino acid change is shown in parentheses. Putative stop codon for met-2Δ is shown with an ‘X’. The SET domain is indicated with light red. b,c, Representative western blot and quantification of H3K9me2 in WT, met-2Δ and met-2-CD (b) or WT, met-2Δ, met-2Δ set-25Δ and met-2-CD set-25Δ (c). N = 3. NS, nonsignificant by two-sided t-test. d, Immunofluorescence and quantification of H3K9me2 normalized to H4 in WT, met-2Δ and met-2-CD. Scale bar, 5 µm. N = 3, n nuclei(embryos): WT = 1,542(49), met-2Δ = 985(51), met-2-CD = 819(39). Median and quartiles shown. NS, not significant by one-way ANOVA, followed by Tukey post hoc test. e, Live imaging of embryos expressing either WT MET-2 or MET-2-CD::mCherry and quantification of MET-2 foci per nucleus. Scale bar, 5 µm. N = 3, nuclei(embryos): met-2::mCherry = 569(30), met-2-CD::mCherry = 613(30). NS, not significant by two-sided Wilcoxon signed-rank test. f, Brood sizes of the indicated strains at 25 °C. N = 2, n = 40. P(WT, met-2Δ) = 4.7 × 10−14, P(WT, met-2-CD) = 0.0002, P(met-2-CD, met-2Δ)=1.04 × 10−13, P(met-2Δ set-25Δ, met-2-CD set-25Δ) = 1.05 × 10−13, by one-way ANOVA, followed by Tukey post hoc test. g, Percentage of synchronized L1s that develop into L4 larvae 42 h after re-feeding at 20 °C. N = 4. P(WT, met-2Δ) = 4.7 × 10−5, P(WT, lin-65Δ;met-2-CD) = 7.2 × 10−4, P(met-2Δ, met-2-CD) = 5.5 × 10−4, P(met-2-CD, lin-65Δ;met-2-CD) = 0.015; ***P < 0.0001, ****P < 0.00001.
