Fig. 8

Involvement of the HBx protein from HBV in AS. a Immunoblotting analysis using anti-HA antibody for the detection of HBx-HA protein from the cell lysates isolated after 5 days of transfection with pLenti6V5A-HBx. Control untransfected cells (T(−)) were also used in this assay. b List of ASEs common to HBV-associated HCC and HBx-expressing cells. c Example of an ASE modified following the expression of HBx. Overview of the two isoforms encoded by ENO3 gene. Exons are depicted in red and the intervening introns are shown as thin black lines (not to scale). The primers used to detect the isoforms by RT-PCR assays are shown in gray and the sizes of the expected amplicons are also indicated (top panel). RT-PCR reactions were performed on control cells (T(−)) and cells expressing HBx (HBx) using specific primers to detect both isoforms of the transcripts encoded by the ENO3 gene. Capillary electrophoresis was performed and an image of the detected reaction products is shown (lower panel). The positions of the expected amplicons are indicated by arrows. d Mass spectrometry analysis of cellular proteins interacting with HBx. The average H/L ratios of the HBx affinity purification-mass spectrometry experiments were plotted versus the total intensities. The red dots indicate cellular proteins involved in RNA splicing