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Fig. 2 The number of immature neutrophils was significantly increased in c-mybhyper;pu.1G242D/G242D zebrafish embryos. (A and B) Accumulation of neutrophils in c-mybhyper;pu.1G242D/G242D embryos. WISH of cebpa, cebp1, and mpx expression at 5 dpf in wildtype, c-mybhyper, pu.1G242D/G242D, and c-mybhyper;pu.1G242D/G242D zebrafish. The caudal hematopoietic tissue (CHT) is enlarged in the red box (200×), n ? 6. (C) qPCR results of 5 dpf embryos from wildtype, c-mybhyper, pu.1G242D/G242D, and c-mybhyper;pu.1G242D/G242D (n >20, three biological replicate). (D?I) The neutrophil maturity and enzyme activity were decreased significantly in the c-mybhyper;pu.1G242D/G242D zebrafish. Granulocyte maturity was detected by SB staining (D), SB+ cell counts (F) and mean optical density (H) were calculated and compared at 5 dpf. The CHT is enlarged in the red box (200×). Green boxes show further enlarged regions focusing on a single cell (400×), n ? 13. Peroxidase activity was detected by DAB staining (E), DAB+ cell counts (G) and mean optical density (I) were calculated and compared at 5 dpf. Red boxes show a further enlarged region (400×), n ? 10.