malbec (mlb^bw306) has normal embryonic and definitive hematopoiesis despite disruption of the cdh5 gene. (A) The expression of erythro-myeloid progenitors (gata1) is normal in wt and mlb embryos at 20 somite stage (ss). wt: n = 80/80; mlb: n = 78/80. (B) WISH analysis of the surrogate marker for myeloid cell expression shows normal mpo levels in mlb embryos at 20 ss. wt: n = 80/80; mlb: n = 75/80. (C) FACS analysis of mpx:eGFP⁺ neutrophils in control and cdh5-silenced zebrafish embryos. Twenty embryos were used in each experiment; n = 5. (D) qRT-PCR analysis of runx1 and c-Myb mRNA expression in control and malbec embryos. Forty embryos were used in each experiment; n = 5. (E) Normal runx1 mRNA expression in the AGM region of 36-hpf-old wt and mlb embryos. wt: n = 220/220; mlb: n = 220/220. (F) A progeny of mlb crossed with c-myb:eGFP has comparable levels of mlb::c-myb:eGFP⁺ HSCs cells in malbec embryos; n = 60. (G) A progeny of mlb crossed with cd41:eGFP has comparable levels of mlb::cd41:eGFP^low HSCs (FACS analysis), indicating that mlb produces sufficient amounts of HSCs. Forty embryos pooled in each sample; n = 5. (H) Normal rag-1 mRNA expression in the thymus of a mlb embryo (examples indicated with arrows). wt: n = 80/80; mlb: n = 80/80. (I) Progeny of mlb crossed with cd41:eGFP shows mlb::cd41:eGFP⁺ platelets (examples indicated with arrows) at day 4 postfertilization. wt: n = 160/160; mlb: n = 156/160. *P < .05 (t test, error bars indicate standard error of the mean [s.e.m.]).