Extended Data Figure 7 : Generation and analysis of RetBCLxL mice.

From: The neurotrophic factor receptor RET drives haematopoietic stem cell survival and function

Extended Data Figure 7

a, Ret locus was targeted by a construct containing the BCLxL coding sequence, an internal ribosomal entry site (IRES) and a puromycin resistance cassette, followed by a floxed neomycin resistance cassette to aid negative selection. Ret Bcl-xL.IRES.Puromicin knock-in mice were obtained by excising of the neomycin cassette. b, Number of myeloid progenitors, LSK cells, multipotent progenitor cells and HSCs in E14.5 fetal liver. WT n = 7; Ret+/BCLxL n = 9; RetBCLxL/BCLxL n = 8. c, FACS-sorted HSCs from Ret+/+, E14.5 fetal liver Ret+/BCLxL and adult bone marrow Ret+/BCLxL were analysed by RT–qPCR for human BCL2L1 expression. Housekeeping genes: Gapdh and Hprt1. d, Annexin V+ cells in cultured E14.5 LSK cells. WT n = 6; Ret+/BCLxL n = 4; RetBCLxL/BCLxL n = 6. e, Scheme of competitive transplantation. RetBCLxL/BCLxL animals and littermate controls were injected in competition with CD45.1/CD45.2 cells, relative to Fig. 4d, e. Error bars, s.e.m. ***P value for one-way ANOVA lower than 0.001.