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. 2014 Mar 10;9(3):e91279. doi: 10.1371/journal.pone.0091279

Figure 4. Impact of ESCRT-II subunits EAP30 and EAP45 on HBV genome maturation.

Figure 4

HuH-7 cells treated with control siRNA (siC) or siRNAs targeting EAP30 or EAP45 were retransfected with the modified pHBVΔHP replicon devoid of foreign promoter elements. A. Lysates were probed with antibodies to EAP30, EAP45, Core, L, and β-actin as indicated. B. Virions released into the cellular supernatants were quantified by real-time PCR of the HBV genomes. Error bars indicate the standard deviations from the mean of two experiments measured in duplicates. C. For quantitative reverse transcription-PCR (qRT-PCR), total mRNAs were isolated, reverse transcribed and used for PCR reactions. To measure HBV pgRNA packaged into intracellular capsids, lysates were subjected to a capsid-specific immunoprecipitation prior to RNA isolation and qRT-PCR. Error bars indicate the standard deviations from the mean of two experiments measured in duplicates.