1, 150 mM NaCl, 1 mM EDTA, 1× complete EDTA-free protease inhibit

1, 150 mM NaCl, 1 mM EDTA, 1× complete EDTA-free protease inhibitors (Roche, Mississauga), 1× phosSTOP phosphatase PI3K inhibitor inhibitors (Roche) and 1% Triton X-100). An equivalent amount of protein from each sample (450 ng) was separated by 10% SDS-PAGE and transferred to PVDF membrane. The membrane was blocked for 1 hour in TBS-T containing 4% BSA, and then incubated in 1:1000 anti-phospho-p44/42 MAPK (Thr202/Tyr204) antibody (#9101, Cell Signaling Technology, Danvers) overnight at 4°C in blocking buffer. The membrane was washed 3× with PBS containing 0.1% Triton X-100, incubated in 1:4000 goat anti-rabbit IgG

HRP-conjugate antibody (Sigma) in blocking buffer for 1 hour at room temperature, washed and developed using enhanced chemiluminescence (ECL) reagents (Amersham, Piscataway). The PVDF membrane was then stripped of antibody, blocked, re-probed with 1:1000 anti-p44/42 MAPK antibody (#9102, Cell Signaling Technology) and developed as above. Transmission Electron Microscopy HeLa cells (1 × 106) in 9 cm2 wells of six-well plates were infected with C. pneumoniae CWL029 at a multiplicity of infection of 1. Compounds were added at 1 hpi and cells harvested at 48 hpi. Cells were fixed overnight at 4°C in 0.1 M sodium cacodylate buffer containing 2% gluteraldehyde, embedded in araldite resin and thin sections were viewed using a Jeol JEM 1200EX electron microscope at 12,000× magnification.

Acknowledgements We thank Dr. Eric Brown and Dr. Gerry Wright for helpful advice and guidance on this project. We are grateful to this website all members of the Mahony lab for stimulating research discussions. A special thanks to Rick McKenzie

for technical help with the Jeol JEM 1200EX electron microscope. Both DLJ and CBS are recipients of a Father Sean O’Sullivan Graduate Scholarship. This work was funded in part by a grant to JBM from the Canadian Institutes of Health Research. References 1. Hahn DL, Azenabor Chlormezanone AA, Beatty WL, Byrne GI:Chlamydia pneumoniae as a respiratory pathogen. Front Biosci 2002, 7:e66-e76.CrossRefPubMed 2. Paldanius M, KU-57788 Juvonen R, Leinonen M, Bloigu A, Silvennoinen-Kassinen S, Saikku P: Asthmatic persons are prone to the persistence of Chlamydia pneumoniae antibodies. Diagn Microbiol Infect Dis 2007, 59:117–122.CrossRefPubMed 3. Sutherland ER, Martin RJ: Asthma and atypical bacterial infection. Chest 2007, 132:1962–1966.CrossRefPubMed 4. Campbell LA, Kuo CC, Grayston JT:Chlamydia pneumoniae and cardiovascular disease. Emerg Infect Dis 1998, 4:571–579.CrossRefPubMed 5. Grayston JT: Background and current knowledge of Chlamydia pneumoniae and atherosclerosis. J Infect Dis 2000,181(Suppl 3):S402-S410.CrossRefPubMed 6. Grayston JT:Chlamydia pneumoniae and atherosclerosis. Clin Infect Dis 2005, 40:1131–1132.CrossRefPubMed 7. Ardeniz O, Gulbahar O, Mete N, Cicek C, Basoglu OK, Sin A, Kokuludag A:Chlamydia pneumoniae arthritis in a patient with common variable immunodeficiency. Ann Allergy Asthma Immunol 2005, 94:504–508.CrossRefPubMed 8.

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