Figure 2.

(A) FACS analysis of MV^D7 cells expressing low (yellow line), medium (red line), and high (light brown line) levels of GFP-tagged Mena and VASP constructs. The black-filled area indicates the background fluorescence of untransfected MV^D7 cells. (B–C′) The expression of wild-type GFP-Mena and GFP-VASP rescues Listeria motility in MV^D7 cells in a concentration-dependent manner. MV^D7 cells expressing high levels of wild-type GFP-Mena or GFP-VASP were infected with Listeria, fixed, and stained with fluorescent phalloidin. In these cells, Listeria recruited GFP-Mena and GFP-VASP at their surface (arrowheads in B′ and C′) and induced the formation of normal actin tails (arrows in B and C; compare with Figure 1). (D) Box and whiskers plots of bacterial speed. Dot indicates the mean, line in the middle of the box indicates the median, top of the box indicates the 75th quartile, whereas the bottom of the box indicates the 25th quartile, and whiskers indicates the 10th and 90th percentiles, respectively. (E–E′) GFP-tagged wild-type Mena and VASP localize at the actin tails induced by the Listeria mutant Δ5. MV^D7 cells expressing high levels of wild-type GFP-Mena were infected with Listeria Δ5, fixed, and stained with fluorescent phalloidin. This Listeria mutant typically induced the formation of short actin tails (arrowhead in E), which were robustly stained with GFP-Mena (arrowhead in E′). Bar (for B–C′ and E–E′), 2 μm.