Figure 6

Decreased proliferation and increased apoptosis of Pim TKO HSCs. (A) Decreased in vivo BrdU incorporation in LT-HSCs of Pim TKO mice. Pim TKO mice or WT mice were injected with 2 doses of BrdU, and BM cells were harvested and labeled with antibodies. BrdU-positive cells in LT-HSCs, ST-HSCs, and MPPs were analyzed (n = 6, **P <0.01). (B) Decreased in vitro Ki67 labeling in Pim TKO LSK cells. LSK cells were sorted from Pim TKO mice or WT mice and cultured in StemSpan medium supplemented with TPO, SCF and Flt3 for 2 days. The cells were then stained with Ki67 antibody. Representative histogram of three independent experiments was shown (MFI: mean florescent intensity). (C) Increased cell death in Pim TKO LSK cells. The cultured LSK cells were stained with Live/Dead dye and representative histogram of three independent experiments was shown. (D) Increased caspase 3-activation in Pim TKO LSK cells. Cultured LSK cells were fixed and stained with caspase 3 antibody. Representative histogram of three independent experiments was shown.