Fig 3

(A) Amino acid sequence is conserved for GTP-binding pockets of the Rab5c proteins between human and zebrafish. The amino acid of zebrafish in red was mutated to generate DN Rab protein by affecting GTP/GDP affinity. (B) Expression of cmyb in control and Rab5c inhibition groups examined by WISH. rab5c DN overexpression was carried out by hsp70-GFP-rab5c DN HS at 20 hpf. Scale bar, 100 μm. (C) Quantification of the cmyb positive cells. Error bars, mean ± SD, ***P < 0.001. (D) WISH analysis shows that cmyb is decreased in embryos of rab5c DN overexpression driven by a fli1a promoter. Scale bar, 100 μm. (E) Quantification of the cmyb positive cells. Error bars, mean ± SD, ***P < 0.001. (F) Endothelial, but not somitic, Rab5c overexpression has a HSPC rescue. Scale bar, 100 μm. (G) Quantification of the runx1 positive cells. Error bars, mean ± SD, ***P < 0.001. The P values in this figure were calculated by Student t test. The underlying data in this figure can be found in S1 Data. DN, dominant-negative; EC, endothelial cell; GDP, guanosine diphosphate; GTP, guanosine triphosphate; HE, hemogenic endothelium; hpf, hours post fertilization; HS, heat shock; HSPC, hematopoietic stem and progenitor cell; n.s., nonsignificant; WISH, whole-mount in situ hybridization; WT, wild type .