FIGURE 3

FIGURE 3

NIPBL‐mediated RUNX1 down‐regulation leads to impaired expression of RUNX1 target genes in both human and zebrafish. A, Spearman's correlation between RUNX1 and MPL in cord blood megakaryocytes (MK) derived from healthy donors. B, Spearman's correlation between RUNX1 and SPI1 in bone marrow cells (BM) derived from 34 adult AML patients without aberrant RUNX1 alterations (mutations or translocations). r = Spearman's correlation coefficient. C and D, RT‐qPCR analyses of 48 hpf ctrl‐, nipblb‐ and nipblb‐MO/runx1mRNA‐injected embryos. C, The expression of the erythroid marker gata1a was decreased following nipblb‐MO injection in comparison with controls and rescued in nipblb‐MO/runx1mRNA‐injected embryos. D, The expression of the myeloid marker spi1b was increased in both nipblb‐MO‐ and nipblb‐MO/runx1mRNA‐injected embryos in comparison with controls. E‐G, O‐dianisidine staining showed a reduction of mature circulating erythrocytes in nipblb‐MO‐injected embryos at 48 hpf in comparison with ctrl‐MO. Co‐injection with the full‐length runx1 mRNA rescues the o‐dianisidine reduction. Lateral views anterior to the left (upper panels) and ventral views of the anterior region (lower panels). H‐J, WISH analyses showed an increased expression of spi1b in nipblb‐MO‐ and nipblb‐MO/runx1mRNA‐injected embryos in comparison with ctrl‐MO. Scale bars indicate 100 μm in (E‐G) and 200 in μm in (H‐J). One‐way ANOVA with Bonferroni correction, ^***P < .001 ^**P < .01, ^*P < .05, n.s: non‐significant