Fig. 4: ARLE-14 stabilizes MET-2 at chromatin redundantly with H3K9me.

a, Brood sizes of the indicated strains at 20 °C. N = 2, n = 40. P(WT, met-2Δ) < 2.0 × 10⁻¹⁶, P(WT, met-2-CD) = not significant, P(WT, arle-14Δ) = not significant, P(WT, arle-14Δ met-2-CD) = 1.94 × 10⁻¹², P(met-2Δ, met-2-CD) = 8.2 × 10⁻¹⁴, by one-way ANOVA, followed by Tukey post hoc test. b–d, Live imaging of expressing either WT MET-2::mCherry or MET-2-CD::mCherry ± arle-14 RNAi (b), and quantification of MET-2 nuclear signal P(WT(vector), WT(arle-14)) = 1.0 × 10⁻¹², P(WT(vector), met-2-CD(vector)) < 2.0 × 10⁻¹⁶, P(WT(vector), met-2-CD(arle-14)) = 5.7 × 10⁻⁷, P(met-2-CD(vector), met-2-CD(arle-14)) < 2.0 × 10⁻¹⁶ (c) and MET-2 foci per nucleus (d). P(met-2-CD(vector), met-2-CD(arle-14 RNAi)) = 0.041. Scale bar, 5 µm. N = 3, n = 45 embryos. Nuclei: met-2::mCherry: vector = 816, arle-14 RNAi = 1,199, met-2-CD::mCherry: vector = 1,056, arle-14 RNAi = 1,787. P values calculated by two-sided Wilcoxon signed-rank test. e, mCherry ChIP-qPCR at representative met-2Δ-specific genes, common genes or the met-2-independent gene unc-119. Bars indicate the mean and dots represent individual measurements. N = 3. Statistical comparison of arle-14Δ met-2-CD with met-2-CD: C55C3.3 P = 0.05, col-166 P = 0.74, bcl-11 P = 0.004, F10D11.6 P = 0.009, grl-16 P = 0.34, glf-1 P = 0.0024, nep-17 P = 0.05, unc-119 P = 0.15, Student’s t-test. f, heatmap of log₂ FC of met-2Δ-specific genes, met-2Δ»met-2-CD genes and common genes in met-2Δ, met-2-CD, arle-14Δ and arle-14Δ met-2-CD relative to WT. g, distribution of log₂ FC of the same comparisons as in (f) displayed as violin plots. met-2Δ-specific: P(met-2Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶, P(met-2-CD, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶, P(arle-14Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶; met-2Δ»met-2-CD: P(met-2Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶, P(met-2-CD, arle-14Δ met-2-CD) = 0.00012, P(arle-14Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶; common: P(met-2Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶, P(met-2-CD, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶, P(arle-14Δ, arle-14Δ met-2-CD) < 2.0 × 10⁻¹⁶ by two-sided Wilcox rank sum test. N = 3. Boxplots show median, boxes 50% and whiskers 90% of each group. NS, not significant; *P < 0.01, **** and #### P < 0.00001.

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