Figure 9.

VipA contributes to membrane wrap formation (A) Representative deconvolved confocal micrographs showing the immuno-labeling pattern depicting the defective barrier formation at membrane wraps formed by dotA and vipA mutants compared to wild-type Lp. LECs were infected for 1 h, washed and fixed 6 h p.i. External bacteria were immunolabeled (red), cells were permeabilized and internal bacteria were immunolabeled using anti- Lp antibodies (blue) and F-actin was stained using phalloidin (green). (B) Quantification from (A). Data shown are mean ± SEM from 3 independent experiments (n>80 filaments in each experiment). (C) Lp attachment to LECs following treatment with cytochalasin-D. Bacterial attachment to vehicle treated controls has been normalized to 100% and attachment following treatment is expressed as relative values. Data shown are mean ± SEM from 3 independent experiments (n>1000 cells in each experiment). For all fluorescence micrographs the main panels are merged z-stacks and the higher magnifications are single z-planes. All scale bars, 12 μm. Statistical analyses were performed using one-way ANOVA with Tukey's multiple comparison test. ^**p < 0.01, ^*p < 0.05.