Figure 6

Binding, trafficking, and degradation of the 16PVs that have bound to DTNB or NEM. (A) The 16PVs were incubated with DTNB(2 mM) or NEM (2 mM) at 37°C for 2 h and added to HeLa cells. After incubation at 4°C for 1 h, the cells were washed by PBS and lysed. The lysate was electrophoresed on an SDS-polyacrylamide gel. L1 was detected by immunoblotting with anti-HPV16L1 antibody. (B) The 16PVs incubated with DTNB or NEM were added to HeLa cells and incubated for 1 h at 4°C. The cells were cultured at 37°C for 2, 4, 8 or 20 h and fixed. L1 was detected by rabbit anti-HPV16L1 antibody and goat anti-rabbit IgG conjugated with Alexa Fluor 546 (red). DNA was stained with DAPI (blue). (C) The 16PVs incubated with DTNB were added to HeLa cells and incubated for 1 h at 4°C. The cells were harvested with PBS containing 2.5 mM EDTA (for trypsin – sample at 0 h) or with trypsin (for trypsin + sample at 0 h). The rest of cells were cultured at 37°C for 2, 4, 8 or 20 h and harvested with trypsin. The cells were lysed and the lysates were electrophoresed on an SDS-polyacrylamide gel. L1 was detected by immunoblotting with anti-HPV16L1 antibody.