TRIM25 was upregulated by IFN treatment in both 293T and HepG2 cells. (a, b) 293T and HepG2 cells were seeded in a 24-well plate. Cells were treated with IFNα (10 ng/ml) as indicated. The treated cells were divided into two groups. One was subjected to immunoblotting using TRIM25 and GAPDH antibody, and the other was subjected to total RNA extraction. The mRNAs were detected using Q-PCR. Data represent the means and s.d. from three independent experiments. Student’s t-test was performed. *P<0.05,**P<0.01. (c) HepG2 cells were treated with IFNα together with CHX (10 μg/ml) or without for 8 h as indicated. Q-PCR and western blotting were performed to analyze the expression of TRIM25. (d) HepG2 cells were IFN-treated for 16 h, and supernatants were collected to add to newly seeded HepG2 cells as indicated. Cells were collected after 8 h, and Q-PCR and western blotting were performed to analyze the expression of TRIM25. Data represent the means and s.d. from three independent experiments. Student’s t-test was performed. *P<0.05, **P<0.01, #P>0.05.