Pre-stained Broad Selleckchem Everolimus Range Protein Markers (New England Biolabs, cat. # 7708) were used for standards. For immunoblotting, separated polypeptides were transferred to Immobilon-P membrane (Millipore), blocked with skim milk (5% [w/v] in Tris buffered saline + Tween20 [TBS-T]) or BSA, and then incubated with specific antibodies at working concentrations; anti-EscJ, 1:500 [69]; anti-EscN, 1/500

[39]; anti-EspB, 1:200 [70]; anti-EspA [71]; anti-intimin 1:1000 (gift from J. Leong); anti-HA, 1:5000 (gift from R. Duncan); anti-FLAG, 1:5000 (Sigma); anti-DnaK, 1:5000 (Calbiochem), anti-Tir, 1:1000 [35]; anti-TEM1, 1:2000 (QED Biosciences); goat anti-mouse conjugated to horse radish peroxidise (HRP), 1:5000 (Rockland immunochemicals); goat anti-rabbit conjugated to HRP, 1:5000 (Rockland immunochemicals); goat anti-rat conjugated to HRP, 1:5000. Anti-CesT polyclonal antibodies were raised in New Zealand white rabbits

against a synthetic peptide (LENEHMKIEEISSSDNK) corresponding to the C-terminal region of CesT (Pacific Immunology, CA, USA, [NIH Animal Welfare Assurance Number: A4182-01]). Final bleeds were affinity purified against the peptide by the supplier and used in immunoblots at a 1:10000 dilution. Immunoblots were developed using an enhanced chemiluminescence reagent (ECL, GE Healthcare) and data captured on a VersaDoc 5000 MP (Bio-Rad). Densitometry measurements to evaluate band intensity from chemiluminescent signals in immunoblotting experiments were GPCR Compound Library concentration performed using Quantity One software (Bio-Rad). Immunoblots were imaged simultaneously and within exposure times that were within an empirically determined linear range of signal detection. Ethics statement The grant proposal supporting this Cetuximab in vitro research was reviewed

by the Dalhousie University Ethics Officer. Ethics approval was not required as the research does not involve human subjects, primary human cell lines/samples or animals. Acknowledgements The authors would like to thank members of the Thomas lab and the Department for critical reading of the manuscript. Madhulika Prasad provided valuable technical assistance with experiments. Roy Duncan graciously provided monoclonal anti-HA antibodies. This research was supported by an operating grant (MOP84472) from the Canadian Institutes of Health Research (CIHR). Infrastructure and research equipment were supported with funds provided by the Canadian Foundation for Innovation Leaders Opportunity Fund (CFI-LOF), the Dalhousie Medical Research Foundation and Dalhousie University. N.A.T is the recipient of a CIHR New Investigator Award. The funding agencies did not participate in study design; in the collection, analysis, and interpretation of data; in the writing of the manuscript; and in the decision to submit the manuscript for publication. References 1. Goosney DL, Celli J, Kenny B, Finlay BB: Enteropathogenic Escherichia coli inhibits phagocytosis. Infect Immun 1999,67(2):490–495.PubMed 2.