FIG 5.

The predicted CPj0678 ORF is expressed in C. pneumoniae-infected cells. (A) Serum samples from eight patients who had been clinically diagnosed with a primary acute C. pneumoniae infection specifically recognize CPj0678-GFP expressed in yeast. Total protein lysates were analyzed by immunoblotting with human antisera or anti-GFP antibodies. The predicted molecular size of CPj0678-GFP is 50 kDa (arrowhead). (B) HEp-2 cells were infected with C. pneumoniae and incubated at 37°C for 72 h. Total proteins lysates were resolved by SDS-PAGE and immunoblotted with antibodies against CPj0678, C. pneumoniae adenylate kinase, or tubulin protein. (C) HEp-2 cells were transfected with CPj0678-GFP for 48 h, fixed, and immunostained with anti-CPj0678 antibodies. (D) HEp-2 cells were infected with C. pneumoniae for the indicated times, and CPj0678 or tubulin protein expression was assessed by immunoblotting with specific antibodies. (E) HEp-2 cells were infected with C. pneumoniae for the indicated time. Cells were fixed and stained with anti-CPj0678 antibodies. C. pneumoniae was detected with FITC-labeled mouse anti-Chlamydia antibodies, and nucleic acids were stained with Hoechst 33342. Images are representative of experiments performed in triplicate.