Journal: Frontiers in Microbiology

Article Title: E2 Site Mutations in S Protein Strongly Affect Hepatitis B Surface Antigen Detection in the Occult Hepatitis B Virus

doi: 10.3389/fmicb.2021.664833

Figure Lengend Snippet: Immunofluorescence staining of HBsAg in HepG2 cells transfected with pHBV1.3B/C and E2 mutated plasmid. The staining of HepG2 cells transfected with pHBV1.3B/C showed a diffuse fine particle staining, while cells transfected with pHBV1.3B/C-E2G/A gathered in the perinuclear region or showed a flaked and coarse grains staining pattern. The staining of cells transfected with pHBV1.3B-E2V/D and pHBV1.3C-E2D was similar to that of genotype-matched pHBV1.3B/C. The cells were fixed at 48 h after transfection and staining with a monoclonal rabbit antibody to the HBsAg, followed by an anti-rabbit IgG conjugated with CY3 (red). The cell nuclei were stained with DAPI (blue). The scale bar stands for 10 μm. Magnification, ×200 or ×400. Mock: pcDNA3.1(+) vector.

Article Snippet: Then the cells were washed three times with 1×PBS before incubation with CY3-conjugated (red) secondary antibodies (goat anti-rabbit) (Affinity Biosciences, China) at room temperature for 1 h. Cell nuclei were stained with DAPI.

Techniques: Immunofluorescence, Staining, Transfection, Plasmid Preparation