Fig. 4

Fig. 4 Gpr171 promotes embryonic HSC development via Notch signaling. (A) Whole-embryo qPCR analysis of Notch signaling gene expression in WT and gpr171−/− embryos at 30 h postfertilization (hpf) (WT, n = 3; gpr171−/−, n = 3; two-tailed, unpaired Student’s t test, *P < 0.05, **P < 0.01. Data are presented as means ± SD). (B) Confocal images of EGFP expression in the dorsal aorta (DA) of WT and gpr171−/− embryos in Tg(tp1:EGFP) background at 30 hpf (white arrowheads). (Scale bar, 100 μm.) (C) Qualification of EGFP+ cells in the DA from images in (B) (WT, n = 14; gpr171−/−, n = 15; two-tailed, unpaired Student’s t test, **P < 0.01. Data are presented as box plot). (D) WISH for cmyb in the VDA of WT and gpr171−/− embryos in Tg(hsp70l:Gal4;UAS:NICD) background at 30 hpf (black arrowheads) after heat shock. (Scale bar, 200 μm.) (E) WISH for cmyb in the VDA at 30 hpf following injection of her15.1 mRNA into WT and gpr171−/− embryos (black arrowheads), and its qualitative phenotypic distribution plot of embryos scored with low, medium, or high cmyb expression. [Scale bar, 200 μm (WT, n = 67; gpr171−/−, n = 61; gpr171−/− + her15.1 mRNA, n = 42).] (F) Confocal images of HSCs in the VDA at 30 hpf following injection of her15.1 mRNA into WT and gpr171−/− embryos in Tg(runx1:EGFP;kdrl:mCherry) background (white arrowheads). Runx1+ cells are in green, and Kdrl+ cells are in red. (Scale bar, 50 μm.) (G) Qualification of Runx1+Kdrl+ cells from images in (F) (WT, n = 14; gpr171−/−, n = 12; gpr171−/− + her15.1 mRNA, n = 15; One-Way ANOVA test, **P < 0.01, ***P < 0.001. Data are presented as box plot).