Fig. 7

H3K27me3 protein level and expression of gata1a and hbbe3 could be recovered effectively by H3K27 methylation inhibitor (EPZ005687) in eaf1^−/− and eaf2^−/− mutants. A Western blotting analysis of H3K27me3 protein level in eaf1^−/−, eaf2^−/− and WT larvae, and the corresponding groups treated with EPZ (EPZ005687) at 24 hpf (A1), and quantification of H3K27me3 protein (A2). B WISH analysis of the expression of gata1a and hbbe3 in eaf1^−/−, eaf2^−/−, WT embryos and the corresponding groups treated with EPZ (EPZ005687) at 24 hpf (B1-B12), and statistical analysis of WISH results (B13, B14). C ChIP-qPCR analysis of the binding enrichment of protein H3K27me3 on the promoter of gene gata1a in eaf1^−/− and eaf2^−/− embryonic cells at both 14 hpf (C1) and 24 hpf (C2), with anti- IgG used as a negative control. D The working model of eaf1 and eaf2 in regulating erythropoiesis. Knockout of eaf1 or eaf2 causes the reduced RBCs and the changed H3K27me3 - gata1a signaling axis, resulting in changes in the binding enrichment of H3K27me3 on the promoter of gene gata1a in zebrafish. Meanwhile, eaf1 and eaf2 promote zebrafish erythropoiesis by modulating the canonical WNT/β-catenin signaling pathway in a developmental stage-specific manner. Each experiment was repeated at least three times, with similar results for two or three replicates, and a representative result are shown. Data are mean ± SD. B1-B12, lateral view, anterior to the left. *P < .05, **P < .01, ***P < .001. NS, not significant. Scale bar = 200 μm (B1-B12)