Fig. 3: CeSPT-2 histone binding is required for transgenerational gene silencing.
From: The histone chaperone SPT2 regulates chromatin structure and function in Metazoa
a, Schematic diagram of transgenerational RNAi; see Extended Data Fig. 3 for the transgenerational RNAi assay described previously41 (figure adapted from ref. 41). b, Representative images of the germline of the gfp::h2b reporter worm strains without RNAi (vector RNAi), after gfp RNAi treatment and in the subsequent generations after removal of the dsRNA (G1–G6). c, Quantification of GFP::H2B expression. Worms showing ‘dim’ GFP intensity during microscopy inspection were considered as silenced. The graph indicates the mean percentage of desilenced worm lines across the replicate lines, and the percentage of desilenced worms in each replicate is indicated by a dot; n = 5. d, gfp::h2b mRNA expression measured by qPCR, normalized relative to cdc-42. n = 1 for ‘empty vector’ datapoint; n = 5 for G1–G6; data are represented as mean ± s.d. Statistical testing: one-way ANOVA, calculated for each generation independently, with Dunnett’s post-tests. NS: P > 0.05; *P < 0.05, **P < 0.01, ***P < 0.0001. One-way ANOVA results, G1: 0.0588, G2: 4.34 × 10−8, G3: 1.19 × 10−4, G4: 4.22 × 10−6, G5: 3.71 × 10−6, G6: 9.75 × 10−8. Exact P values of Dunnett’s post-test comparisons shown in the figure are reported in Supplementary Table 3. e, Representative microscopy images of the germline of the indicated worm strains without RNAi (vector RNAi), after gfp RNAi treatment and in the subsequent generations after removal of the dsRNA (G1–G9). f, Quantification of GFP::H2B expression, as in c. The graph indicates the mean percentage of desilenced worm lines across the replicate lines, and the percentage of desilenced worms in each replicate is indicated by a dot; n = 5 (WT and hrde-1 mutant); n = 3 (for each of the spt-2HBM siblings).
