Fig 1. siRNA-knockdown screening for ESCRT factors required for HBV replication in HepG2 cells.

(A) A cartoon illustrates the ESCRT cascade and the interactions between each component members of ESCRT-0, I, II, III, and the VPS4 ATPase complex. Of note, the exact stoichiometry of certain component members remains tentative [5]. (B) An HBV replicon plasmid containing a genomic dimer and individual siRNA against each ESCRT factor were co-transfected into HepG2 cells. Viral DNA replication, viral RNA expression and intracellular capsid particles were analyzed by Southern, Northern and Western blot analysis at day 5 post-transfection. The 28S rRNA was included as an RNA loading control. RC: relaxed circular DNA, DL: double-strand linear DNA, SS: single-strand DNA. We detected significant effect on HBV replication from 18 out of 30 tested ESCRT factors (as summarized in Table 1). Data shown here are representative of at least three independent experiments.