Figure 3.

Inhibition of mTORC1 protects against palmitate-induced cell death in hepatocytes. AML12 cells were pretreated with either torin-1 (0.25 µM) or rapamycin (0.25 µM) for 2 h before palmitate (0.4 mM) exposure. Cell death was determined after 16-h palmitate exposure via both LDH release assay (a) and MTT measurement (b); (c) AML12 cells were pretreated with Triacsin C (10 µM), the long-chain fatty acyl-CoA synthetase inhibitor, for 2 h prior to palmitate exposure (0.4 mM). Cell death was determined 16-h later by LDH release; (d) AML12 cells were pretreated with GSK1940029 (20 µM), the stearoyl-CoA desaturase (SCD)-1 inhibitor, for 2 h before palmitate addition (0.4 mM). Cell death was determined after 16-h palmitate exposure by LDH release measurement. (e) AML12 cells were pretreated with PF-4708671 (10 nM), the S6K1 inhibitor, for 2 h before palmitate addition (0.4 mM). Cell death was determined after 16-h palmitate exposure by LDH release measurement. AML12 cells were pretreated with SC79 (10 µM), an Akt activator, for 2 h before palmitate addition (0.4 mM). Akt activation status and cell death were detected by Western blotting for p-Akt protein abundance (f) and LDH release measurement (g) 16 h later, respectively. All values are denoted as means ± SD from three or more independent experiments. Bars with different characters differ significantly (P < . 05).