Figure 1.

HBV minichromosome establishment occurs very rapidly after infection. (A and B) Southern blot analysis of cccDNA appearance kinetic. PHHs or HepG2^hNTCP cells were infected with HBV in the presence or not of 100 nmol/L preS1-mimicking peptide for up to 16 hours and then harvested at the indicated times points. Mitochondrial DNA was used as an internal loading control. The specificity of the cccDNA band is shown by linearization after digestion with EcoRI or XhoI restriction enzymes. (C–H) qPCR quantification of viral cccDNA, 3.5-kb RNA, and total HBV DNA (tHBV-DNA) in (C–E) PHHs and (F–H) HepG2^hNTCP-infected cells. Cells were inoculated with HBV for up to 16 hours and then harvested at the indicated time points. cccDNA and tHBV-DNA quantification were normalized over β-globin quantity, while the relative 3.5-kb RNA amount was normalized over the housekeeping gene GUSb expression. Graphs represent the means ± SEM of at least 3 independent experiments. MW, molecular weight; p.i., post infection.