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. 2015 Apr;100(4):431–438. doi: 10.3324/haematol.2014.106468

Figure 2.

Figure 2.

Analysis of hematopoietic stem and progenitor cells (HSPCs) in Cdk2fl/flCdk4−/−vavCre (DKO) mice. Flow cytometric analysis of HSPCs in Cdk2fl/flCdk4−/−vavCre DKO mice (red bars) and wild-type or double heterozygote Cdk2+/flCdk4+/−vavCre littermate controls (blue bars). (A) Bone marrow (BM) frequency of CD71hiTER119lo pro-erythroblasts (pro-E) (n=4). (B) BM frequency of TER119hi CD71hiFSChi basophilic erythroblast (EryB), TER119hi CD71hi FSClo polychromatic erythroblast (EryP) and TER119hiCD71loFSClo orthochromatic erythroblast (EryO) (n=4). (C) Frequency of c-Kit+Sca-1+Lineage− (KSL) hematopoietic stem/progenitor cell (HSPC) compartment in the BM (n=5). (D) Flow cytometry analysis of Ki-67 expression in KSL cells. Percentage of KSL cells in the various cell cycle phases is shown (n=4). (E) BM frequencies of individual HSPC compartments (n=6). LT-HSC, ST-HSC and MPP compartments are gated from viable c-Kit+Sca-1+Lineage− (KSL) cells. CMP, GMP and MEP compartments are gated from viable c-Kit+Sca-1−Lineage− cells. CLP compartment are gated from viable c-KitlowSca-1lowLineage− cells. LT-HSC: long-term hematopoietic stem cell; ST-HSC: short-term hematopoietic stem cell; MPP: multipotent progenitor; CMP: common myeloid progenitor; GMP: granulocyte/macrophage progenitor; MEP: megakaryocyte/erythroid progenitor; CLP: common lymphoid progenitor. (F) BM frequencies of indicated lineage cells (n=4). Myeloid: Mac-1+ and Gr-1+; B cells: B220+ CD19+; natural killer (NK) cells: CD3-Dx5+; megakaryocyte (MgK): CD41+CD61+. (G) Frequency of indicated lineage cells in the thymus (n=6). DP: double positive T cell; SP: single positive T cell. (H and I) Bone marrow samples from Cdk2fl/flCdk4−/− vavCre mice (DKO) or from littermate controls (Control) were labeled with BrdU in culture for the indicated time periods (n=3). (H) The percentage of BrdU+ CD71+ erythroblasts (labeling index) are indicated. (I) The plot of BrdU labeling index of CD71+ bone marrow erythroblasts versus the cumulative BrdU labeling time is shown. Error bars represent standard deviation. Two-tailed t-test results are indicated by asterisks; *P<0.05, **P<0.01, ***P<0.001.