Leflunomide alleviates lipid accumulation in FFA-stimulated AML12 cells via the AMPK pathway. (A-B) AML12 cells were transfected with control siRNA (si-control) or AMPKα siRNA (si-AMPKα) for 24 h, and then treated with or without leflunomide (100 μM) or teriflunomide (100 μM) for 30 min. The representative immunoblots of p-AMPKα, AMPKα, p-ACC, and ACC were shown and the p-ACC/ACC ratio was calculated. n = 5. (C-E) AML12 cells were first transfected with si-control or si-AMPKα for 24 h and then were treated with FFA and leflunomide (100 μM) or teriflunomide (100 μM) simultaneously for 18 h. (C) The Nile red stain was performed and the respective images were shown. Scale bar = 20 μm. (D-E) The representative immunoblots of p-AMPKα, AMPKα, p-ACC, and ACC were shown, and the p-ACC/ACC ratio was calculated. n = 5. (F) Representative immunofluorescent images of SREBP1 (green) and the cell nucleus stained with DAPI (blue). n = 5. One-way ANOVA analysis was performed: *P < 0.05, ** P < 0.01, *** P < 0.001.