Figure 2

The dynamic changes of HBV transcripts, intracellular viral DNA and cccDNA expression profiles in different cell growth phases. 1.3ES2 cells (2.5 × 10⁶) were initially plated onto a 10 cm diameter Petri dish and cultured continuously for 24 days. The culture medium was refreshed every three days during the experimental period. (A) Total RNA was extracted at the indicated time points and subjected to Northern blot analysis to determine the expression profiles of the 3.5-kb, 2.4-kb, and 2.1-kb HBV transcripts. A GAPDH-specific probe was used to control for equal sample loading. (B) Total DNA was extracted at the indicated time points, digested with HindIII, and then separated by electrophoresis. The expression profiles of the integrated viral genome, RC DNA, and DL DNA were examined by Southern blot analysis. The integrated viral genome was used to control for equal sample loading. (C) The intracellular cccDNA was collected by Hirt extraction at each time point. The cccDNA was thermally denatured at 85°C for 5 min, and then subjected to Eco RI digestion before electrophoresis on a 1.2% agarose gel and Southern blot analysis.