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The EMBO Journal (1998) 17, 2767–2776, doi:10.1093/emboj/17.10.2767 Figure 5 Neural Wiskott–Aldrich syndrome protein is implicated in the actin-based motility of Shigella flexneri Toshihiko Suzuki, Hiroaki Miki, Tadaomi Takenawa and Chihiro Sasakawa

Figure 5 Actin assembly by Shigella is inhibited by transient expression of dominant-negative N-WASP in Cos-7 cells. (A) Cell lysates from Cos-7 cells expressing N-WASP variants were used for a binding assay with GST–1. Immunoblots of whole cell lysates are shown in the upper panel. Immunoblots of N-WASP bound to GST–1 beads are shown in the lower panel. (B) Quantitation of the actin-associated intracellular bacteria in transfected cells. The transfected cells were infected with S.flexneri (M94 carrying pD10-1) or L.monocytogenes 1/2a EGD. The data shown are the means of triplicate experiments. The top bars show the standard deviation of the mean. (C) Cos-7 cells overexpressing full-length N-WASP (a, c and e) or cof mutant (b, d and f) were infected with Shigella and immunostained using anti-N-WASP antibody (a and b) and rhodamine–phalloidin (c and d). The yellow color in the combined image shown in (e) and (f) indicates co-localization between N-WASP (green) and F-actin (red). Bars, 10 m.

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