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. 2004 Jan 5;199(1):35–46. doi: 10.1084/jem.20030204

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Copyright © 2004, The Rockefeller University Press

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Figure 5. — The ITAM-like sequence of CEACAM3 is critical for bacterial internalization and Rac stimulation. (A) Primary sequence of the COOH-terminal cytoplasmic domain of CEACAM3 containing the ITAM-like sequence. The ITAM consensus (31) is shown below with the critical tyrosine residues in bold. (B) 293 cells were transfected with the control vector (pcDNA) or the indicated HA-tagged CEACAM3 constructs. Whole cell lysates (WCL) were analyzed with anti–HA tag antibody. (C) Transfected cells as in B were infected with Opa_CEA-expressing gonococci for 60 min and analyzed in gentamicin protection assays. The graph shows mean values ± SDs of three to four independent experiments done in triplicate. (D) Cells were transfected as in B, and samples were lysed before or after 60 min of infection with Opa_CEA N. gonorrhoeae. GST-CRIB pull-down assays (top) or whole cell lysates (WCL; bottom) of the samples were analyzed with monoclonal anti-Rac antibody.