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. 2024 Jul 2;20(10):3725–3741. doi: 10.7150/ijbs.93465

Figure 5.

Figure 5

The effects of leflunomide and teriflunomide on lipid accumulation and AMPK activation in AML12 cells were mediated by inhibiting DHODH. (A-B) The AML12 cells were first transfected with empty vector pCDNA3.1 or plasmid pCDNA3.1-mDHODH for 24h, and then cells were treated with leflunomide (100 μM) or teriflunomide (100 μM) for 30 min. (C-F) After transfected with empty vector pCDNA3.1 or plasmid pCDNA3.1-mDHODH for 24 h, the AML12 cells were treated with FFA (500 μM) and leflunomide (100 μM) or teriflunomide (100 μM) simultaneously for 18 h. (A-B & D-E) The representative immunoblots of p-AMPKα, AMPKα, p-ACC, and ACC were shown and the ratio of p-ACC/ACC and p-AMPKα/AMPKα were analyzed and calculated. n = 5. (C) Nile red staining was performed and the respective images were shown. Scale bar = 20 μm. (F) The immunofluorescent staining was performed with specific SREBP1 antibody (green) and DAPI (blue) for cell nucleus staining. One-way ANOVA analysis was performed: *P < 0.05, ** P < 0.01, *** P < 0.001.