pY14Cav1 regulates FA turnover and cellular displacement in MDA-435 human breast cancer cells. (A) MDA-435 cells were transfected with FAK-GFP alone or FAK-GFP with either Cav1-mRFP or CavY14F-mRFP and FAK-GFP imaged every 30 s for 30 min. Representative images at time 0, 6, 12, 18, 24, and 30 min are shown (see Videos 1–3). (B) Overlay of FAK-GFP images at time 0 (red) and 30 min (green) shows FA displacement over time (top) and cell outlines show cellular displacement over time (bottom). Protrusive regions are indicated by green arrows and retractile areas by red arrows. (C) Stable, disassembled, and newly formed FAs were quantified as a percentage of total FAs during the 30-min interval of acquisition. (D) Retractile and protrusive cell areas over the 30-min interval of acquisition and total cell area at 30 min were normalized to cell area at time 0 and displayed as a percentage (mean ± SEM; n = 4; *, P < 0.05; **, P < 0.01 relative to FAK-GFP only). Bar, 20 μm.