To test steatosis prevention, treatment with ACC1/2 inhibitors Firsocostat (6.0 μM, 0.6 μM, and 0.06 μM), PF-05175157 (5.0 μM, 0.5 μM, and 0.05 μM), and MK-4074 (0.3 μM, 0.03 μM, 0.003 μM) was begun 72 h prior to treatment with FFA and continued for an additional 4 days (Fig. 3A). This evidence concerns the gene ACACA and steatosis.