Expression levels of transcription factors associated with the regulation of HBV transcription. (A) Total RNA was extracted at the same time points and subjected to Northern blot analysis to determine the expression profiles of the HBV transcripts: the 3.5-kb, 2.4-kb, and 2.1-kb transcripts (upper panel). The amount of sample loaded was normalized with a GAPDH-specific probe (lower panel). (B) The expression of transcription factors that regulate HBV pgRNA transcription was assessed by quantitative real-time RT-PCR at the indicated time points. The bar chart shows the relative expression of HNF4α, HNF1α, HNF3β, CEBPα, PPARα, RXRα, HLF, NR2F1, and NFIL3 mRNAs which was normalized to that of B2M. ■ day 3 sample; ■ day 6 sample; ■ day 9 sample; ■ day 12 sample. * P < 0.05, ** P < 0.01, and *** P < 0.001. The data were analyzed statistically with Student's t test. (C) The expression profiles of hepatocyte-specific markers (albumin, TAT, G6P mRNAs) were determined by real-time RT-PCR. The results are shown after normalization to B2M. ■ day 3 sample; ■ day 6 sample; ■ day 9 sample; ■ day 12 sample. (D) The culture media were collected at the indicated time points, and the amounts of secreted albumin were determined by ELISA.