Effect of Y-27632 treatment on FA reorganization in HeLa JW cells.

<div><p>(<b>A</b>) Localization of GFP-tagged VASP, zyxin, paxillin, vinculin, talin, kindlin-2, ILK and FAK in untreated cells (control) or in the cells treated with 10 µM Y-27632 for 10, 30 or 90 min (shown are snapshots from time-lapse movies). White and black arrowheads point to peripheral and central adhesions, respectively.</p> <p>(<b>B</b>) FA lifespan in three different cells tagged by each of the tested proteins prior to and during Y-27632 treatment, as tracked by time-lapse video microscopy. Each line marks the lifespan of an individual adhesion site, from its formation until its complete disassembly. Individual cells are marked by asterisks on the Y axes. The vertical dashed line marks the beginning of the Y-27632 treatment. Peripheral and central FAs were analyzed separately. Note the decay of existing FAs and formation of novel FAs during Y-27632 treatment.</p> <p>(<b>C</b>) Western blots showing MLC and pMLC in untreated cells, or in cells treated with 10 µM Y-27632 for 10, 30 and 90 min Tubulin was used here as a loading control.</p>