Figure 5.
HDAC11 is expressed in mouse BM cell subsets and Hdac11 loss does not interfere with normal hematopoiesis. (A) Schematic of mouse models and experimental design used to assess the role of HDAC11 in normal hematopoiesis. (B-D) eGFP-Hdac11 expression in Tg-Hdac11-eGfp reporter mice53 in BM subpopulation as determined by flow cytometry. (E) Absolute stem and progenitor cell counts from Hdac11−/− and matched Hdac11+/+ from 1 tibia and 1 femur as determined by flow cytometry. The data represent 3 independent experiments with 6 mice per genotype per experiment. (F-H) WBC, platelet (PLT), and RBC counts from Hdac11+/+(n = 5), Hdac11+/− (n = 9), and Hdac11−/− (n = 9) mice (8 weeks old) are shown. (I) In vitro colony formation counts from Hdac11+/+ or Hdac11−/− WBM cells (1 × 104) seeded in methylcellulose media for 7 days. The data represent 2 independent experiments with 2 to 3 mice per genotype per experiment. (J-N) BM cells (1 × 106) from CD45.2 Hdac11+/+ (n = 4) or Hdac11−/− (n = 4) mice transplanted into lethally irradiated recipient mice (n = 15 recipients per genotype) along with CD45.1 recipient BM cells (1 × 106). Donor cell reconstitution in the myeloid (Mac1+ cells), B-cell (B220+), and T-cell (CD3+) lineages from peripheral blood for 2 to 10 weeks after transplantation. (O) BM chimerism and absolute donor cell numbers of lineage-negative (Lin–), LSK, HSPCs (LT-HSCs and ST-HSCs) 10 weeks after transplantation. Flow gating for this experiment and for LT-HSCs and ST-HSCs is shown in supplemental Figure 5B-C). Additional methods for antibodies and instrument settings are provided in supplemental Tables 1 and 2. Additional supporting data are provided in supplemental Figure 5D-L. CLP, committed lymphoid progenitor; CMP, committed myeloid progenitor. Statistical analysis was performed by using multiple two-tailed unpaired Student t test. ****P < .0001; ***P < .001; **P < .01; *P < .05.