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. 2004 Jul 19;166(2):225–235. doi: 10.1083/jcb.200402073

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Copyright © 2004, The Rockefeller University Press

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Figure 8. — Coupled action of Crk and cortactin is required for efficient uptake of Shigella. HeLa cells were transfected with vector alone (mock), FL cortactin (FL), Myc-tagged wild-type Crk (wtCrk), or HA-tagged mutant Crk (R38VCrk), or were cotransfected with Myc-tagged wild-type Crk and Flag-tagged FL cortactin (wtCrk-FL), HA-tagged mutant Crk, and Flag-tagged FL cortactin (R38VCrk-FL). After 24 h of transfection, cells were infected with Shigella, fixed, and processed for fluorescent labeling. (a) The frequency of Shigella-induced actin foci formation at 15 min was scored for each transfectant. (b) Shigella uptake was determined for each transfectant using outside/total staining. The results of each experiment were normalized to the value obtained for mock transfectants, corresponding to 37% uptake. The average number of cell-associated bacteria was 4.5 bacteria per cell, and did not vary significantly in the different samples. The plotted data were averaged from the scoring of samples from three independent experiments ± SEM. Statistical significance was assessed by t test (denoted by asterisks).