Host cell-dependent late entry step as determinant of hepatitis B virus infection
Fig 5
HEK293 cells were rendered susceptible to HBV and HDV infections after huNTCP expression.
(A) Western blot analysis of deglycosylated cell lysates from parental and huNTCP-expressing HEK293 cells for validating huNTCP expression. β-actin was used as the loading control. (B) SDS-PAGE and western blot analysis of whole cell lysates of parental and huNTCP-expressing cells at 8 days post-infection with HDV for detecting HDAg using the human HDAg antibody. (C) SDS-PAGE and western blot analysis of whole cell lysates of mock-infected or HDV-infected huNTCP-expressing cells were harvested at 2-, 4-, 6-, and 8-days post-infection with the human HDAg antibody. (D) Parental and huNTCP-expressing HEK293 cells were plated on regular culture dishes (i.e., with no collagen coating) and infected with ca. 400 genome equivalent of HBV per cell. Three days post infection, the PF DNA (i.e., Hirt DNA) from mock- or HBV-infected cells was extracted by Hirt extraction and treated with Exo I/III followed by Southern blot analysis. Hirt DNA from HBV-infected HepG2-huNTCP cells, loaded at 4-fold less than the Hirt DNA from HEK293 and HEK-293-huNTCP cells, served as the positive control for cccDNA detection.