Figure 5

Vinculin recruitment to cell-cell contacts occurs with F-actin stabilization in MDCK cells. (A) Confluent MDCK cell monolayers on collagen-coated coverslips were incubated with media containing 200 nM jasplakinolide (Jas) or media alone (Control), fixed, and immuno-stained for vinculin and F-actin (phalloidin) at cell-cell contacts. The same exposure and laser power were used to acquire and generate the images. Scale bar 10 μm. (B) Data of immuno-fluorescence intensities and analysis of F-actin and vinculin at cell-cell contacts are presented as mean ± standard deviation and are compared with control (n = 10, ***P < 0.001). (C) Single cell ablations of confluent MDCK wild-type cell monolayers were performed following treatment with media containing 200 nM Jas, 50 μM blebbistatin (Bleb), or media alone (Control). Vinculin and actin accumulated along the wound edge in Jas-treated and control cells, but was reduced with Bleb treatment. Scale bar 10 μm. (D) Comparison of the initial velocity of wound-closure between control (n = 8) and Jas-treated (n = 7) cells. Error bars are standard error of the mean.