Effect of constitutive expression of mutated IDH1 in primary bone marrow cells in vitro and in vivo. (A) Cumulative CFC yield is shown for an initial plating of 1000 transgene-expressing cells (cells were sorted before plating; mean ± SEM of 3 independent experiments). (B) Protein expression of mouse bone marrow cells transduced with a CTL vector or with FLAG-tagged wild-type IDH1 or mutant IDH1 using an anti-FLAG antibody. Cells were harvested from bone marrow 4 weeks after transplantation. β-actin was probed on the same blot as a loading control. (C) Ratio of R-2HG to S-2HG in mouse bone marrow cells transduced with CTL vector or with wild-type or mutant IDH1. Cells were harvested from bone marrow 4 weeks after transplantation (mean ± standard error of the mean [SEM] of 3 independent mice). (D) Engraftment of transduced cells in peripheral blood at different time points after transplantation of 1 × 10⁵ transgene-positive cells (mean ± SEM). (E) White blood cell count in peripheral blood at different time points after transplantation of control or IDH1-transduced bone marrow cells (mean ± SEM). (F) Hemoglobin levels in peripheral blood at different time points after transplantation of control or IDH1-transduced bone marrow cells (mean ± SEM). (G) Platelet count in peripheral blood at different time points after transplantation of control or IDH1-transduced bone marrow cells (mean ± SEM). (H) Immunophenotype of transduced cells in peripheral blood at 16 weeks after transplantation (mean ± SEM). (I) Average spleen weight in mice receiving transplants of control or IDH1-transduced bone marrow cells 20 weeks after transplantation (mean ± SEM). *P < .05; **P < .01; ns, not significant. WB, western blot.