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. 2009 Nov 11;106(47):19975–19979. doi: 10.1073/pnas.0908365106

Fig. 2.

Fig. 2.

The viral transactivator HBx is recruited onto the cccDNA in HBV replicating cells. (A, Upper) Cross-linked chromatin from HepG2 cells transfected with monomeric linear full-length HBV DNA was immunoprecipitated with the relevant control IgG or specific anti-HBx, anti-AcH3, and anti-AcH4 antibodies and analyzed by PCR with HBV cccDNA selective primers. (Lower) ChIPed samples were analyzed by PCR using primers specific for the cyclin A2 coding region as a negative control. (B) Kinetics of HBx recruitment onto the HBV cccDNA. (Upper) ChIP analysis was performed on chromatin from HepG2 cells cotransfected with monomeric linear full-length WT HBV DNA and an HA-tagged HBx expression vector using the relevant control IgG or specific anti-HA tag and anti-AcH3 specific antibodies. (Lower) Exogenously expressed HBx is detected by anti-HA immunoblotting. Tubulin levels detected by immunoblotting were used to normalize equal loadings from lysate samples.