HBx modulated expressions of target proteins in transiently transfected and HBV producing malignant hepatocytes. (A) Western blot confirmed proteins PTEN, p27 and MAP3K (Raf 1) – targets of miR-21, miR-222 and miR-145 respectively were increased accordingly in HBx transfected HepG2 cells. Cells were transfected with 1 μg and 2 μg of HBx plasmid respectively or pCXN2 as a control plasmid. (B) Expressions of PTEN, p27 and MAP3K (Raf 1) proteins in 1.3 fold HBV transfected HepG2 cells. HepG2 cells were transfected with 1 μg pUC19-HBV or 1 μg pUC19 vector as a control. (C) Western blot exhibited differential expression of target proteins PTEN, p27 and MAP3K (Raf 1) in constitutively HBV producing HepG2.2.15 and control HepG2 cell line. Cells were collected for protein analysis 48 h after each transfection. β actin was taken as endogenous control. Protein bands were quantified using dentiometric scanner (Bio-Rad). Below are the graphical representation of intensity ratio between target proteins (PTEN, p27 and MAP3K (Raf 1)) and β actin in each lane.