Figure 6.

HBV-Mediated NF-κB Activation Transcriptionally Regulates the Expression of TRIM37

(A–D) LX-2 cells were transfected with HBV replicon or control vector, cells were treated with NAC (10 mM) for 12 h, and (A) ROS production was measured by dichlorofluorescin diacetate (DCF-DA) staining followed by fluorescence-activated cell sorting (FACS), and the fluorescence intensity was quantified; (B–D) expression levels of TRIM37 and NF-κB p65 were measured by RT-PCR and/or western blotting. (E and F) LX-2 cells were transduced with either control vector or HBV replicon, treated with PDTC (10 μM) or vehicle for 24 h, and (E) luciferease reporter activity of TRIM37 promoter was measured, and (F) mRNA and protein levels of TRIM37 were measured by RT-PCR and western blotting, respectively. (G) The NF-κB binding site in the the TRIM37 promoter was predicted by the JASPAR algorithm. (Left) The predicted sequence, binding site (BS), and schematic diagram of TRIM37 gene, promoter region, and 3′ UTR are shown. (Right) Chromatin from LX-2 cells was immunoprecipitated using control IgG or NF-κB p65 antibody, and binding of the antibody in the BS region of the promoter or negative control 3′ UTR region of TRIM37 (NC) was measured by qPCR. ∗∗∗p < 0.001 compared with control or IgG; ^###p < 0.001 compared with HBV.