Fig. 6

Gfi1aa expression is lost in definitive haematopoietic cells of homozygous qmc551 fish, but definitive haematopoiesis is normal. (A,C,F-O) Fixed embryos stained in WISH experiments using indicated probes. Whole embryos are shown in (A,H,J,M-O). Close-up views of anterior and posterior parts of the embryos are presented in (G) and (C,F,I,K), respectively. All embryos, except (G), are shown in a lateral view with anterior to the left and dorsal up. (G) shows a close-up dorsal view with anterior to the left. (L) shows a transverse section through the trunk of one of the homozygous qmc551 embryos after gfi1ab WISH. (B,R) QRT-PCR on total RNA isolated from whole 26 hpf embryos (B) and from adult KM cells (R) to measure the relative level of gfi1aa mRNA (Two-tailed t-test: p=0.002 (26 h), p=0.034 (KM)). Ef1α mRNA was used as a loading control. (D,E) Live images of qmc551 embryos with anterior left and dorsal up. Close-up pictures of the tail (D) and the head regions (E) are presented. (P) Regulation of Gfi1aa and Gfi1ab expression at the onset of definitive haematopoiesis. (Q) Flow cytometric analysis of green and red fluorescence in adult KM cells excited with a 488 nm laser and detected using 529/28 and 580/23 nm band pass filters, respectively. Please note that a substantial proportion of KM cells displays green and red autofluorescence. (S) May-Grünwald/Giemsa stained KM cytospins. (T) Relative number of bilobed and trilobed neutrophil granulocytes observed in May-Grünwald/Giemsa stained cytospins. (U) Sudan Black staining on adult KM cytospins. Arrows: red - prRBCs, green - HECs and definitive HCs, yellow - inner ear hair cells, white - T cell precursors in the thymus, black - neutrophil granulocytes with bilobed nuclei. Arrowheads: green - reduced gene expression in the vDA.