Cdh5 is dispensable for HSC development in E11.5 AGM and E13.5 fetal liver and reconstitution of multilineage adult hematopoiesis. (A) Quantitative flow analysis of CD41, c-Kit, and CD45.2-positive cells in E11.5 Cdh5^−/−GFP⁺ and WT (Cdh5^+/+GFP⁻) cells, indicating that loss of Cdh5 is dispensable for CD41, c-Kit, and CD45.2 expression. Eleven AGM were used for FACS analyses. (B) CFU analysis of sorted E11.5 AGM-derived WT and Cdh5^−/− GFP⁺ cells, indicating that loss of Cdh5 has no influence on hematopoietic colony differentiation. Twelve AGM were used for CFU analyses; 2 e.e. per 3.5 mL of M3434 media. (C) Photographs of green fluorescent GFP⁺ hematopoietic colonies formed from the E11.5 AGM-derived Cdh5^−/−GFP⁺ HSCs. Magnification, ×4. (D) Schema showing the transplant of 2 e.e. E11.5 whole chimeric AGM in irradiated SJL mice. (E) Quantitative analysis of peripheral blood from mice transplanted with 2 e.e. E11.5 chimeric whole AGM, indicating that Cdh5^−/−GFP⁺ HSCs reconstitute to multilineage adult blood up to 16 weeks of transplant. Twelve AGM were transplanted in 6 SJL recipients. (F) Quantitative analysis of the percentage of Lineage⁻Sca1⁺c-Kit⁺ (LSK) cells in WT (Cdh5^+/+GFP⁻) and Cdh5^−/−GFP⁺ compartments of E13.5 fetal liver, indicating that loss of Cdh5 has no intrinsic influence on development of LSK HSCs. Six fetal livers were used for FACS analyses. (G) CFU analysis of sorted E13.5 fetal liver–derived sorted WT (Cdh5^+/+GFP⁻) and Cdh5^−/−GFP⁺ cells, indicating that loss of Cdh5 has no influence on hematopoietic colony differentiation. Sorted cells from 10 fetal liver were used for CFU analyses. (H) Schema showing the transplant of E13.5 fetal liver–derived GFP⁺ or GFP⁻ sorted cells in irradiated SJL mice. (I) Quantitative analysis of peripheral blood from mice transplanted with E13.5 fetal liver–derived WT and Cdh5^−/−GFP⁺ sorted cells, indicating that Cdh5^−/−GFP⁺ HSCs reconstitute multilineage adult blood up to 16 weeks of transplant; n = 6 recipients. (J) Percentage GFP⁺CD45.2⁺ peripheral blood chimerism from the Cdh5 knockout cells in E11.5 AGM and E13.5 fetal liver–transplanted recipients. Each dot represents an individual recipient; n = 6; *P < .05 (t test, error bars indicate s.e.m.).