HBV Virions Separation from Culture Supernatant

Detailed experimental procedures of HBV virions isolation were described in our previous report (Liu et al. 2020). Briefly, HBV virions were immunoprecipitated from culture supernatant by anti-HBs antibody conjugated to protein G beads (#11243233001; Roche) overnight at 4 °C. In control groups, samples were incubated with beads only. For virion associated HBV DNA detection, the precipitate was digested with RQ1 RNase-Free DNase (#M6101, Promega, Madison, WI, United States) and Plasmid-safe DNase I (#E3101K; Epicentre Inc., San Diego, CA, United States) according to the manufacturer’s protocol. The viral DNA was further extracted and detected by Southern blot and real time-PCR.