Fig. 3

H3K27me3 epigenome editing by dCas9-olEzh2(∆SET) and higher concentration injection. a–c dCas9-olEzh2(∆SET) analyses targeting Arhgap35 promoter. ChIP-qPCR using anti-FLAG antibody (a), ChIP-qPCR using anti-H3K27me3 antibody (b) and Arhgap35 mRNA expression fold change (c). d–e Arhgap35 mRNA expression fold change (d) and ChIP-qPCR using anti-H3K27me3 antibody (e) in higher concentration injection. f The epigenetic modification patterns around Kita, sgRNAs (blue bars) and ChIP-qPCR product (black bars) positions. H3K27me3 (red) and H3K27ac (blue) ChIP-seq [27], DNase I-seq (black) [28] and DNA methylation enrichment [34] at the blastula stage are shown for comparison. g–i dCas9-olEzh2(∆SET) analyses targeting Kita promoter. ChIP-qPCR using anti-FLAG antibody (g), ChIP-qPCR using anti-H3K27me3 antibody (h) and Kita mRNA expression fold change (i). In Fig. 4c, e and i, after expression levels were normalized to that of beta-actin, fold changes (sample/no injection) were calculated. Light blue, gray, orange, red and pink bars in each bar graph represent no injection, sgRNAs/dCas9 injection, sgRNAs/dCas9-olEzh2 injection, sgRNAs/dCas9-olEzh2(∆SET)(350 nM) injection and sgRNAs/dCas9-olEzh2(∆SET)(550 nM) injection, respectively. (Tukey–Kramer test, *p < 0.1, **p < 0.05, ***p < 0.01, n = 3 biological replicates and only in c, d and i n = 6 biological replicates, error bars are s.d., p values of each comparison are shown only if the p value is under 0.1.)