FIG 5.

Replication and protein expression of recombinant NCCR BKPyV. (A) RPTECs were transfected with the indicated recombinant BKPyV NCCR genomes. Supernatant viral loads were quantified by quantitative PCR after 1, 2, 3, 5, and 7 days posttransfection (dpt). Increases in BKPyV supernatant viral loads are expressed on a log scale. (B) Immunofluorescence was performed for large T antigen (LTag; red), agnoprotein (agno; green), and viral protein 1 (VP1; cyan) at 7 dpt. The nuclei are marked with DAPI (blue). The graph shows expression of the indicated viral proteins relative to that of the archetype ww(1.4) protein. (C) Western blot of indicated cell extracts showing LTag (triangles) and VP1 in green (triangles); actin (red) served as a loading control. Actin-normalized band fluorescence intensity (bottom diagrams) was quantified using Licor Image Studio, with ww(1.4) as a reference.