Figure 2.

Stable HBx transfection results in transformation of L-O2 cells. (A) Western blot analysis showed that AFP was detectable in human hepatoma H7402 cells (positive control), but not L-O2 cells transiently transfected with HBx (L-O2+HBx), suggesting that transient overexpression of HBx could not transform L-O2 cells. β-actin was used as loading control (lower panel). (B) Western blot analysis showed that AFP could be detected in both H7402 and L-O2-X cells, suggesting that stable HBx transfection promotes transformation of L-O2 cells. The histogram shows the results after analysis with the Glyco Band-Scan software. Results shown are representative of three independent experiments. (C) Representative photographs of soft agar colony formation were taken three weeks after culture of engineered cells. The diagram shows the mean colony counts from three petri dishes (error bars indicate SEM, n=3 experiments, ^cP<0.01, Student's t test). (D) Number of nude mice with tumor formation (n=8 per group) 3 weeks after injection with engineered cells. Only 3 of 8 mice injected L-O2-X cells generated tumors. Western blot analysis showed the expression levels of HBx and AFP in the tumor tissues. β-actin was used as loading control (lower panel). The histological analysis revealed the pathological features of the tumors (H&E. staining).