Absence of p65 alters the differentiation of HSPCs. (A) Bone marrow or (B) spleens of p65hem−/− mice or wild-type controls were analyzed for the frequency of lineage-committed cells by flow cytometry (n ≥ 7). (C) 5 × 106 whole bone marrow cells from p65hem−/− mice or littermate controls were transplanted into lethally irradiated recipients. Twenty weeks after transplant, bone marrow was harvested, and the frequency of lineage-committed cells was analyzed by flow cytometry (top; n = 5). All recipients displayed >98% engraftment by CD45.2 cells. (D) Bone marrow was analyzed for the expression of p65 by immunoblotting. (E) One hundred LSKF cells were cultured in methylcellulose media with cytokines for 7 days. The average number of colonies per well is displayed. Data are representative of 3 separate experiments. *P < .005. (F) A complete blood panel was performed on peripheral blood collected via cardiac puncture from p65hem−/− and wild-type counterparts (n ≥ 7).
