Fig. 6.

HBx promotes cell proliferation via UCP. We transduced or not parental or F-HBx-expressing HepG2 cells with Ad-F-UCP or Ad-LacZ at a MOI of 50, incubated them for 24 h, and analyzed them by WB as indicated (a). Cell cycle determined using Muse Cell Analyzer in the parental or F-HBx-expressing HepG2 cells with Ad-F-UCP or Ad-LacZ at an MOI of 50 (b). c We transduced or not parental or F-HBx-expressing HepG2 cells with Ad-shUCP or Ad-shControl at an MOI of 200, incubated them for 24 h, treated with BrdU, and cell viability was then determined for 3 days. We repeated this assay twice, each in triplicate. Data are mean ± SD. d–f We transduced or not parental or F-HBx-expressing HepG2 cells with Ad-shUCP or Ad-shControl at an MOI of 200, incubated them for 24 h, and injected them (2 × 10⁷ cells/mouse) into nude mice (N = 5 per group) subcutaneously. After cell inoculation, tumor growth was monitored by measuring tumor volume with digital caliper at the indicated times (d). Mice were killed at day 24, and tumors were then excised from mice and photographed (e). HBx–UCP–VHL–HIF axis was analyzed by western blotting in tumor xenografts (f). We repeated the assay of d–f twice; representative data are shown. Immunohistochemistry was performed with HBx, UCP, and Ki-67 antibodies on paraffin-embedded tumor xenografts (g). The relative intensity of DAB staining was quantified using Image J software, and graphed (h). (i.e., *p < 0.05, **p < 0.01)