Role of mTORC1 and mTORC2 in survival and metabolic activity of mature huMCs and HMC-1.2 MCs. (A) HuMCs were cultured in the presence of vehicle alone, mTORC1/mTORC2 (Torin1, 200nM) or mTORC1 (rapamycin, 200nM) inhibitors and caspases were analyzed by immunoblotting. As a positive control, cell samples from UV-irradiated cells cultured for 8 hours were used. (B) HuMCs were cultured as in panel A for 5 days and cell viability was determined by annexin V assay. As a positive control, imatinib (2μM) was used. (C) HuMCs were cultured as in panel A and metabolic activity of the cells was determined by the MTT assay and evaluated. (D-F) HMC-1.2 MCs were investigated as for huMCs in panels A through C, with the exception that dasatinib treatment in panel E was performed for 3 days. (G-J) HuMCs or HMC-1.2 MCs were cultured as in panel A and the numbers (G and I) and cell viability (H and J) were evaluated. In panels A and D, the blots are representative of 3 donors (A) or 3 independent sample preparations (D). In panels B, C, and E through J, data represent means and SEM (n = 3; 3 donors or 3 independent sample preparations) and differences among the differentially treated samples of each MC type are indicated (*P < .05 by 1-way ANOVA).