MTA1 promoter activity in HepG2 cells (A) or HEK 293 (B) transfected with either control vector
(250 ng/reaction) or HBx expression vector (250 ng/reaction) from different expression systems
(pcDNA 3.1, pSI-GFP, pSG5) was analyzed. Full-length murine MTA1 promoter (■)(-140 to -
5200) or its minimal promoter (-2872-5200)(□) were cloned into pGCL3 luciferase reporter vector
(Promega, Madison, WI). Luciferase assay were performed according to the manufacturer's
instructions (Promega, Madison, WI), and the results were normalized against the -galactosidase
activity, an internal control. (C) Western blot analysis for transfection efficiency in HepG2 after
being cotransfected with pcDNA-p65 expression vector and HBx expression vector. Transfected
cells were analyzed for HBx, NF-κB-p65 48 hours after transfection. Vinculin was used as a
control. (D) NF-κB promoter activity in HepG2 cells transfected with either control vector or HBx
expression vector or HBx-deletion constructs (∆C-HBx-HBx with intact 101-120 amino acids and
∆E-HBx-HBx with intact 101-120 amino acids).
qPCR analysis of MTA1 and HBx (WT and mutant) mRNAs in HepG2 cells transfected with either
control vector or HBx or mut-HBx expression vector. Expression levels of analyzed mRNAs were
normalized by β-Actin.