Figure 2. Immunophenotyping of bone marrow and peripheral blood cells in Hspa9^+/− mice are normal compared to Hspa9^+/+ littermates.

A) No difference was observed in bone marrow (left panel) or peripheral blood (right panel) of Hspa9^+/− and Hspa9^+/+ littermates analyzed by flow cytometry for immunophenotypic markers for (bars labeled top to bottom) neutrophils (Gr1⁺/CD115⁻), B-cells (B220⁺), monocytes (Gr1^lo/CD115⁺) and T-cells (CD3e⁺) (N=3–6/genotype at each time point). B) Red blood cell precursors evaluated by flow cytometry (CD71^+/−/Ter119⁺) in bone marrow of Hspa9^+/− and Hspa9^+/+ littermates at 12 months of age showing no difference between genotypes. C) Bone marrow cells from 12-month-old Hspa9^+/− and wild-type littermates were stained with immunophenotypic markers for SLAM (Lin⁻Sca⁺cKit⁺CD150⁺CD48⁻), KLS (cKit⁺, Lin⁻, Sca⁺), megakaryocyte-erythrocyte progenitors (MEP, Lin⁻Sca⁻cKit⁺FcγR^loCD34⁻), granulocyte-monocyte progenitors (GMP, Lin⁻Sca⁻cKit⁺FcγR^hiCD34⁺), and common myeloid progenitors (CMP, Lin⁻Sca⁻cKit⁺FcγR^loCD34⁺). Bone marrow cells were isolated at indicated time points from Hspa9^+/+ and Hspa9^+/− mice, D) plated in CFU-C media (10,000 cells/plate) and counted on day 7 or E) plated in mature BFU-E media containing only erythropoietin (100,000 cells/plate) and counted on day 10–11. (Hspa9^+/+, filled circles; Hspa9^+/−, open circles) ProEBs, proerythroblasts; BasoEBs, basophilic erythroblasts; PolyEBs, polychromatic erythroblasts; OrthoEBs, orthochromatic erythroblasts. Statistical analysis by two tailed Student’s t-test. Error bars represent mean ± SD. *p<0.05