Role of mTORC1 and mTORC2 in the growth of CD34⁺ cells, mature huMCs, and LAD2 and HMC-1.2 MCs. (A) One-week-old cultures of huCD34⁺ progenitors were supplemented with vehicle alone (Ctrl), mTORC1/mTORC2 (Torin1, 200nM), or mTORC1 (rapamycin, 200nM) inhibitor, cultured for 72 hours, and the effect of the drugs on cell numbers was evaluated. (B) One million huMCs were incubated as in panel A and the numbers of cells in the cultures were determined and evaluated. (C) One million LAD2 MCs were incubated as in panel A, and the number of cells in the cultures was determined. (D) One million HMC-1.2 MCs were incubated as in panel A, and the number of cells in the cultures was determined. (E-F) HMC-1.2 MCs (E) or huMCs (F) were supplemented with vehicle alone (Ctrl) or rapamycin (200nM), cultured for 5 days, and the effects on raptor and rictor protein levels were evaluated. In panels A through F, the data represent means and SEM (A-B, n = 3, and F, n = 2, where n represents the number of donors; C-D, n = 3, and E, n = 6, where n represents the number of independent sample preparations) and differences among the differentially treated samples (*P < .05; for C-D by 1-way ANOVA and for E-F, by t test) are indicated.