Depletion of primitive macrophages does not affect HSPC mobilization and colonization of the CHT. (A-D) WISH of mfap4 shows macrophages are absent in cebpa^rj31 mutant compared with control sibling. White arrows indicate mfap4-positive cells in the yolk sac. (E-H) WISH of cmyb shows macrophage-deficient cebpa^rj31 mutants have normal levels of HSPC in the AGM and the CHT, respectively. (E'-H') Magnified images of corresponding boxed regions from panels E-H, respectively. (I-J) Fluorescent images of the CHT region of cebpa^rj31/Tg(cd41:GFP) embryo at 72 hpf. (K) ImageJ analysis revealed no significant difference in the number of cd41:GFP⁺ cells in the CHT at 72 hpf. Two-tailed Student t test. (L) Flow cytometry analysis showed no significant changes in the relative number of HSPC/cd41:GFP^low+ cells between cebpa^rj31 mutants and control siblings (0.756% ± 0.237% vs 0.599% ± 0.041%, P > .05, 2-tailed Student t test). The data are representative of 3 independent experiments. FSC-H, forward scatter height; GFP-A, green fluorescent protein area; hpf, hours postfertilization; n.s., not significant.