Long-term repopulation deficiency is not caused by altered cell cycle or survival. (A) Cell-cycle analysis (Hoechst 33342) of E12.5 FL LSKs from Wt and Wnt3a^−/− embryos. Values represent percentage of LSK cells in S/G2/M phases of the cell cycle. Results are representative of 3 Wnt3a^−/− and 6 Wt embryos from 3 different litters. (B) Determination of percentage of apoptotic cells by staining with annexin V and 7AAD. Apoptotic cells were defined as annexin V⁺ 7AAD⁻. Results are representative of 6 Wnt3a^−/− and 6 Wt embryos from 2 different litters. (C) Activation of Notch1 in E12.5 FL LSKs from Wt and Wnt3a^−/− embryos. Results are representative of 4 Wnt3a^−/− and 8 Wt embryos belonging to 4 different litters.