Figure 3

Figure 3

HSPC expansion and maintenance in the CHT are compromised in prpf31^−/−zebrafish.A–I and A’–I’, time-course analysis of the expression of HSPC markers runx1 and cmyb in prpf31^−/− and sibling embryos from 28 hpf to 48 hpf by WISH. The reduced expression of cmyb in prpf31^−/− zebrafish was rescued by prpf31 mRNA injection at 48 hpf. Lateral views. Red arrows denote the AGM region. Black arrows denote the CHT region. The number of embryos with similar gene expression patterns among all embryos examined was shown at the bottom right of each panel. The scale bars represent 250 μm. J, quantification of mRNA signals of prpf31^−/− zebrafish and siblings detected by WISH in (A–I and A’–I’). The total number of embryos examined was indicated below each column. Mean ± SD; unpaired two-tailed t test; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; ns, not significant. K, in vivo observation of HSPCs using transgenic line Tg (cmyb: EGFP) of prpf31^−/− zebrafish and siblings, in the AGM and CHT at 36 hpf, 48 hpf, and 3 dpf. The total number of embryos examined was indicated at the bottom right of each panel. The scale bars represent 50 μm. L, quantification of the number of HSPCs in the AGM and CHT of prpf31^−/− zebrafish and siblings observed in (K). The total number of embryos examined was indicated below each column. Mean ± SD; unpaired two-tailed t test; ∗p < 0.05, ∗∗∗p < 0.001; ns, not significant. AGM, aorta-gonad-mesonephros; CHT, caudal hematopoietic tissue; dpf, days postfertilization; hpf, hours postfertilization; HSPC, hematopoietic stem and progenitor cell; PRPF, pre-mRNA processing factor; WISH, whole-mount in situ hybridization. EGFP, enhanced green fluorescent protein.