Overexpression of Rheb2 decreased long-term competitive repopulation in vivo. (A) PB chimerism of transduced GFP⁺ donor (CD45.2⁺) cells and total donor (CD45.2⁺) cells in the 1:1 transplantation group (2 independent experiments, at least 11 animals in each group; mean ± SEM). (B) PB chimerism of transduced GFP⁺ donor (CD45.2⁺) cells and total donor (CD45.2⁺) cells in the 0.4:1 transplantation group (2 independent experiments, at least 11 animals in each group; mean ± SEM). (C) Primary recipient BM chimerism of transduced GFP⁺ donor (CD45.2⁺) cells and total donor (CD45.2⁺) cells in 1:1 group from one of the experiments in panel A (6 Mieg3 and 5 Rheb2 animals; mean ± SEM). (D) PB chimerism of transduced GFP⁺ donor (CD45.2⁺) cells and total donor (CD45.2⁺) cells in secondary transplantation recipients (4 secondary recipients per primary animal; mean ± SEM). *P < .05. **P < .02. NS indicates not significant. (E) Real-time PCR analysis of p21, PTEN, and Gfi-1 gene expression in transduced 32D cells. HPRT was used as internal control to standardize expression of all genes in both cell lines. Data are expressed as fold change over levels in 32D-Mieg3 cells, p21 and Gfi-1 are combined results from 3 experiments, each done at least in duplicate, whereas PTEN is combined results from 2 experiments, each done at least in duplicate. *P < .05. **P < .02.