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. 1999 Aug 31;96(18):10176–10181. doi: 10.1073/pnas.96.18.10176

Figure 6.

Figure 6

Role of T-plastin in S. typhimurium entry into cultured cells. COS-1 cells were transfected with plasmids encoding HA-epitope-tagged wild-type human T-plastin (C, ○), its dominant-negative form (A and C, ⊞), or the same vector encoding the green fluorescent protein (B and C, ▴). Transfected cells were infected with wild-type S. typhimurium with a multiplicity of infection of 50 (C). Data presented in C are averages of two independent experiments. In each experiment, a minimum of 300 COS-1 cells was counted in every category at each time point. Recruitment of plastin to the S. typhimurium-induced membrane ruffles. COS-1 cells were transfected with a plasmid encoding an HA-epitope-tagged human T-plastin either alone (D–F) or together with a plasmid expressing CDC42HsN17 (5-fold excess) (G). Transfected cells were infected with wild-type S. typhimurium for 20 minutes (D, E, and G) or left uninfected (F), and cells were fixed and stained with a monoclonal antibody directed to the HA epitope tag to visualize T-plastin (green, D, F, and G), rhodamine phalloidin to visualize F-actin (E), and 4′,6′-diamidino-2-phenylindole (DAPI) (E) or rhodamine-labeled anti S. typhimurium O antigen antibody to stain bacteria (G).