Irf2bp2a gene is repressed by Gfi1aa.(A) Luciferase reporter assay. Bars showed the relative luciferase activity on the zebrafish irf2bp2a promoter (-2.1 kb) (Student t test, N = 3. Error bars represent mean ± SEM. ***P < 0.001). (B) CHIP-PCR analysis of irf2bp2a promoter in zebrafish embryos expressing GFP, Gfi1aa-GFP or Gfi1aa-Δzinc finger-GFP using an anti-GFP antibody. The statistical significance was calculated by using one-way ANOVA. The asterisk indicates a statistical difference (N = 4. Error bars represent mean ± SEM. ns: not statistically significant, ****P < 0.0001). (C) irf2bp2a relative expression level analyzed by quantitative PCR in GFP-positive cells sorted from control Tg(mpx:eGFP) embryos and gfi1aa MO injected Tg(mpx:eGFP) morphants at 2 dpf (Student t test, N = 5. Error bars represent mean ± SEM. ****P < 0.0001).
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