Figure 3. Long-lived wdr-5 mutants have more H3K9me2 enrichment than wild type.

(A) Immunoblot comparing H3K9me2 protein levels in late-gen wild type to late-gen wdr-5 mutants and late-gen met-2 mutants (representative of two independent experiments). Actin is used as a loading control. (B, D, E) Metaplots of averaged z-score H3K9me2 ChIP-seq signal across H3K9me2 peaks (B), all genes (D), or germline genes (E) in early-, mid-, and late-gen populations of wild type (yellow, orange, and red, respectively) and wdr-5 mutants (lavender, purple, and plum, respectively). Line shows mean ChIP-seq signal. Data in (B) are also shown separated into wild type (B’) and wdr-5 mutants (B’’). Plots are centered on peak centers (B, B’, B’’) or pseudoscaled over genes to 1 kb with 500 bp borders on either side, indicated by vertical gray lines (D–E). (C) H3K9me2 ChIP-seq ratios of wdr-5 mutant coverage over wild-type coverage at each H3K9me2 peak (N = 10,457). Coverage is normalized to RPM. Thick line shows mean and whiskers show standard deviation. Green dots represent peaks that fall beyond y-axis scale (two peaks in mid-gen and 33 peaks in late-gen). ****p<0.0001 with paired t-test.

Figure 3—figure supplement 1. Long-lived wdr-5 mutants have more H3K9me2 than wild type.

(A)-(B) Representative H3K9me2 enrichment profiles in an IGV browser screenshot over a 26 kb region of Chr. III (A) and the rsks-1 locus (B). Average z-scored ChIP-seq tracks for H3K9me2 signal in early-gen wild type (yellow), early-gen wdr-5 mutants (lavender), mid-gen wild type (orange), mid-gen wdr-5 mutants (purple), late-gen wild type (red), and late-gen wdr-5 mutants (plum). All coverage tracks are shown on the same scale, −1.1 to 6.2 (A) or −1.5 to 3.5 (B). Below the tracks, H3K9me2 ChIP-seq peaks are shown in green, and genes are shown in blue. Highlighted region outlined in green shows a peak where mutant/wild type coverage ratios do not change over generational time. Highlighted regions outlined in pink show peaks where the ratio increases in late-gen populations. (C) Overlap between peaks called in transgenerational ChIP-seq experiments of wdr-5 mutants (Figure 3) or jhdm-1 mutants (Figure 4).