Fig. 4

p53 mRNA and protein expression in HBx transfected HepG2 and HepG2.2.15 cells after RNAi treatment. a Relative expression of p53 in HepG2 cells after transfection with HBx expressing plasmid pCXN2-HBx or control vector. Cells are transfected with 1 μg of HBx plasmid or pCXN2 as a control. RNA were collected 48 h after each transfection. b Relative expression of p53 in HepG2 cells by co-transfection with HBx plasmid (0.8 μg) and HBx mRNA specific siRNA (40 p mole) or Negative Control. RNA were collected 48 h after each transfection. c HepG2.2.15 cells were transfected with HBx siRNA (80 p mole) or Negative Control and relative expressions of p53 was measured. RNA were collected 24 h post transfection. Data plotted are the mean ± SD normalized to GAPDH expression. The experiments were performed in triplicate (*P < 0.05, **P < 0.01, ***P < 0.001; Student’s t-test). d Corresponding p53 protein expression was analysed by western blot in HepG2 cells transfected with 1 μg of HBx plasmid or empty vector as a control. e p53 protein expression was analysed in HBx silenced HepG2 cells transfected with HBx plasmid. HepG2 cells were co-transfected with HBx plasmid (0.8 μg) and HBx mRNA specific siRNA (40 p mole) or Negative Control f Relative Expression of p53 protein was measured in HepG2.2.15 cells after siRNA treatment. HepG2.2.15 cells were transfected with HBx siRNA (80 p mole) or Negative Control. Cells were collected for protein analysis 48 h after each transfection. For HepG2.2.1.5, cells were collected at 24 h post transfection. β actin was taken as endogenous control. Protein bands were quantified using Densitometric scanner (Bio-Rad). Below are the graphical representations of intensity ratio between p53 and β actin in each lane