Figure 2

Expression of CT223 at different times post infection and differential reactivity with specific antibodies. DNA in all panels is labeled with DAPI (blue) and the bar in panel F represents 10 microns in each image. Cells were infected at an MOI of approximately 0.2 and fixed with 100% methanol prior to antibody labeling. Panels A-D: Fluorescent microscopy of McCoy cells infected with either strain LGV-434 (A, B) or J/UW36 (C, D). Cells were fixed at different times p.i. (A: 12 h, C: 18 h, B, D: 38 h). In panels A-D, cells were labeled with monoclonal anti-CT223p antibody (green) and anti-HSP60 (red). Note that labeling of CT223p is patchy in each strain at the early times points p.i. (A, C) but the labeling is distinct between strains at 38 h p.i. (B, D). Panels E-H: Cells were infected with strain J/UW36 (E, F) or J(s)1980 (G, H) and fixed 30 hours p.i. Cells were then labeled with either polyclonal anti-CT223p antisera (E, G) or monoclonal anti-CT223p antibody (F, H), both of which are labeled red. Note that CT223p is labeled by the polyclonal antisera in each strain, while the monoclonal anti-CT223p does not label the protein in strain J(s)1980.