Fig. 6

H3K4me3 regulates mlh1 and brip1 expression to maintain CHT HSPC development. (A) Western blotting (Left) and statistical data (Right) showing the protein level of Brip1 and Mlh1 in the CHT region of siblings and wdr5 mutants at 2.5 dpf. Error bar, mean ± SD, **P ≤ 0.01. (B) Genome browser view showing the enrichment of H3K4me3 at putative regulatory elements of mlh1 and brip1 in the CD41+gata1− HSPCs of sibling and wdr5 mutants. (C) H3K4me3 CUT&Tag qPCR showing the relative enrichment levels of H3K4me3 at the mlh1 and brip1 promoter in HSPCs in the CHT region of siblings and wdr5 mutants at 2.5 dpf. Error bar, mean ± SD, *P ≤ 0.05. (D) WISH (Left) and statistical data (Right) showing the expression of cmyb (black arrowheads) in brip1 siblings, brip1 mutants, mlh1 siblings, and mlh1 mutants. Error bar, mean ± SD, ****P ≤ 0.0001. (Scale bar, 100 μm). (E) WISH (Left) and statistical data (Right) showing the expression of cmyb (black arrowheads) in siblings, wdr5 mutants, sibling×Tg(hsp:flag-mlh1-EGFP; hsp:myc-brip1-mCherry) and wdr5 mutant×Tg(hsp:flag-mlh1-EGFP; hsp:myc-brip1-mCherry). mlh1 and brip1 driven by hsp70 promoter were induced by heat shock at 28 hpf. Error bar, mean ± SD, n.s.: no significance, *P ≤ 0.05, ****P ≤ 0.0001. (Scale bar, 100 μm).