Fig. 2.

Production and characterisation of ΔHBc HBV.

(A) Huh7 cells were either transfected with a WT HBV construct (pHBV1.1) or co-transfected with a construct containing a HBV 1.1 mer with stop mutation (lightning bolt) in the HBc ORF (pHBV1.1-HBc^stop) along with a construct expressing WT HBc (pHBc). Supernatant from transfected cells was collected and purified by heparin affinity chromatography. (B) Precipitated supernatant was loaded on caesium chloride density gradients and fractions were analysed by dot blot. Naked capsids (NC) and virions (VP) were identified by the density of the fractions. (C) HepG2-NTCP cells were infected with WT or ΔHBc HBV at increasing inoculating dose. Supernatant was collected from 5 to 7 days post inoculation (dpi) and extracellular HBsAg and HBeAg were measured by ELISA. MyrB-mediated blockade of HBV infection was used as a negative control. The mean (line) of 2 independent experiments (circles/squares) is shown. (D) The numbers of HBs and HBc-positive HepG2-NTCP cells were determined by immunofluorescence 7 dpi. Cells infected at mge 200 are shown. Scale bars represent 100 μm (main figure) and 20 μm (inset). HBV, hepatitis B virus; mge, multiplicity of genomic equivalent; NTCP, sodium taurocholate cotransporting polypeptide; WT, wild-type.