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Fig. 7. Localization of myosin VIIa and uptake of InlA-coated beads and Listeria in cells expressing truncated forms of vezatin. (A) Schematic representation of vezatin variants fused to GFP. The transmembrane fragment (TM) of full-length vezatin (VezFL) is indicated in black. The region containing the myosin-VIIa-interacting site is indicated. Dotted lines indicate regions absent from each construct. (B) Expression level of each vezatin construct in L2071 hEcad fibroblasts. The fluorescence intensity of GFP was quantified in 150 transfected cells (50 cells per coverslip) for each vezatin-GFP construct and compared with that of untransfected cells. Values are means±SD of three experiments. (C) Localization by fluorescence microscopy of myosin VIIa and vezatin-GFP fusions in L2071 hEcad cells expressing the VezFL-GFP, VezC-ter and VezC-terT constructs. Arrows show sites where myosin VIIa and vezatin-GFP constructs are recruited around entering InlA-coated beads. (D) Proportion of intracellular beads and bacteria compared with the total number of associated beads or bacteria in L2071 hEcad cells expressing the different vezatin constructs. Values are means±SD of three experiments, each done in triplicate.

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