Figure 4

Reduction of Mcl-1 is involved in pro-apoptotic effect of HBx in response to oxidative stress. (A) HepG2 cells were infected with Ad-Mcl-1, Ad-shMcl-1 or Ad-blank (M.O.I.= 5) for 36 hr and protein levels of Mcl-1 and GAPDH were determined by Western blot assay. (B, C and D) Cells were infected with indicated adenoviruses (M.O.I. = 5) for 36 hr followed by exposed to H₂O₂ (400 μM) for additional 15 hr. Expression of PARP, caspase-3, Mcl-1, Bcl-xL, HBx and GAPDH was evaluated by Western Blot assay. "ns", nonspecific bands. (E) HBx-Tg mice were subjected to a single injection of Mcl-1-expressing plasmid (p3×flag-Mcl-1) or control plasmid (p3×flag) by tail vein using a hydrodynamics-based procedure (see "Materials and Methods"). Three days later, mice were challenged with liver I/R. Livers were collected and subjected to TUNEL and H.E. staining. Representative results are shown. Magnification, ×100. (F) Mice were treated as in Fig. 4E and circulating levels of serum ALT and AST were analyzed. Results are mean ± SD (n = 4). *, p < 0.05.