1. Furin-dependent infection following ECM attachment.
Furin-dependent infection following ECM attachment. (A) A titration series of HPV16 pseudovirus containing a packaged GFP expression plasmid was applied to MCF10A-derived ECM in conditioned medium from FD11 cells or CHOΔfurin cells (indicated as −/+ furin, respectively). Unbound virus and medium were removed by washing, the indicated target cells were added, and infection was allowed to proceed for 2 days. Pseudovirus infection was determined by flow cytometric analysis of GFP+ cells. The HeLa transfer condition shows the infection derived from unbound pseudovirus that was transferred to empty wells. HeLa cells were then added, and infection was allowed to proceed as described above. (B) A titration series of HPV16 pseudovirus containing a packaged GFP expression plasmid was applied to the matrices indicated in conditioned medium from either FD11 cells or CHOΔfurin cells (indicated as −/+ furin). Unbound virus and medium were removed by washing, pgsa-745 target cells were added, and infection was allowed to proceed for 2 days. Pseudovirus infection was determined by flow cytometric analysis of GFP+ cells. (C) A titration series of HPV16 pseudovirus containing a packaged GFP expression plasmid was applied to MCF10A-derived ECM in conditioned medium from CHOΔfurin cells for the indicated time. Unbound virus and medium were removed by washing, the target cells were added, and infection was allowed to proceed for 2 to 3 days, depending upon the time of virus removal. The target cells were either pgsa-745 (P), HeLa (H), or HeLa and 5 μM furin inhibitor (H+I). Pseudovirus infection was determined by flow cytometric analysis of GFP+ cells.
Patricia M. Day et al. Clin. Vaccine Immunol. 2012; doi: /CVI