Figure 9

Investigation of colocalization of ECTV-PH and VACV-ATI by immunofluorescence. HA-tagged VACV-ATI protein, a truncated-type ATI protein, and Myc-tagged ECTV-PH were overexpressed in BS-C-1 cells using a vTF7-3 transient expression system. (A) Control cells, infected with vTF7-3 and transfected with calf thymus DNA, show DAPI staining of cellular nuclei and little background staining with anti-HA and anti-Myc antibodies (negative control). (B) DAPI staining of cellular nuclei and viral DNA. Discrete areas of DNA in the cytoplasm are indicated by arrows 1, 2 and 4. (C) VACV-ATI protein (green) and (D) ECTV-PH (red) are both distributed throughout the cytoplasm. Arrows 1, 2 and 3 indicate areas of high protein colocalization. (E) Merged view of panels (B-D). Arrows 1 and 2 indicate putative inclusion bodies where VACV-ATI, ECTV-PH, and viral DNA colocalize. Arrow 3 indicates the colocalization of VACV-ATI and ECTV-PH to a putative protrusion from the cell surface.