Overexpression of Ezh2 in HSCs. (A) WBC counts of mice given transplants with BM cells transduced with control () or Ezh2 () vectors (n = 11 recipients per group, 2 independent experiments). Mean values plus or minus standard error of the mean (SEM) are shown. (B) Chimerism at different time points after transplantation. The percentage of donor-derived transduced CD45.1⁺GFP⁺ WBCs is shown. Average values ± 1 SEM of 2 independent experiments are shown. (C) Ezh2 protein expression in the spleen about 120 days after primary transplantation of control or Ezh2 CD45.1 BM cells. (D) CAFC frequencies of sorted LSK GFP⁺ cells 120 days after primary transplantation (control, ; Ezh2, ). (E) Chimerism levels of secondary recipients, competitively transplanted with various ratios (2:1, ⋄; 1:1, ○; 1:2, □; 1:5, ▵) of transduced/nontransduced and freshly isolated BM cells, analyzed 3 months after transplantation. Values show data from individual recipients in 2 independent experiments (n = 35/group). (F) CRI calculated for CD45.1⁺GFP⁺ (transduced) versus CD45.1⁺GFP^- (nontransduced) cells (see insert). Values (+ 1 SEM) are averages 3 months after secondary transplantation, based on 11 and 24 individual mice in the first and second experiment, respectively (control, ; Ezh2, ). (G) CRI calculated for transduced cells (CD45.1⁺GFP⁺) compared to freshly isolated BM cells (CD45.2⁺; see insert). Averages values (+ 1 SEM) of 11 and 24 individual mice of the first and second experiment, respectively, are shown (control, ; Ezh2, ). (H) LTRA frequencies in CD45.1⁺GFP⁺ (transduced) cell fractions calculated from limiting dilution analyses 3 months after secondary transplantation in 2 independent experiments (control, ; Ezh2, ).