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Figure 5. Clustering results in tyrosine phosphorylation of TirMC and recruitment of Nck, N-WASP, Arp2/3 complex, and α-actinin. (A) HeLa cells were either mock transfected and infected with wild-type EPEC (top panels) or were transfected with plasmids encoding TirMC or TirMC(Y474F) and challenged with E. coli expressing intimin (middle and bottom panels). Cells were stained with antibodies to detect phosphotyrosine (Y-PO4), Nck, N-WASP, Arp3, or α-actinin (all shown in green). Bacterial DNA was visualized by DAPI staining (blue). (B) HeLa cells were mock transfected and infected with EPECΔtir + pTir (left lane), or were transfected with a plasmid encoding TirMC and treated with particles coated with anti-TirM antibodies (right lane). HA-tagged Tir was immunoprecipitated from cell lysates and Western blotted for HA-Tir and phosphotyrosine. Image published in: Campellone KG et al. (2004) Copyright © 2004, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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