Autophagy promotes FA turnover in migrating cells. (A) Spinning disk confocal microscopy time-lapse sequences of migrating cells expressing paxillin-mCherry (black) to monitor FA dynamics. Left panels show representative cells expressing shCTRL (top), shATG7 (middle), or shATG12 (bottom). Image sequences of boxed regions have been rotated such that the cell edge with dynamic FAs is moving upward vertically. Elapsed time (min) shown in top left. Bars, 5 µm. Insets are magnified twofold. These images correspond to Videos 2 and 3. (B) Example plots of paxillin-mCherry fluorescence intensity (y axis) over time (x axis) for shCTRL (left), shATG7 (middle), and shATG12 (right) cells used for calculating FA turnover parameters in C. Plots generated by manually tracking individual FAs over time, and each data point is a three-frame running mean of intensity value. The green line represents FA assembly fitted with a logistic function, and the red line represents FA disassembly fitted with an exponential decay function. The lifetime is the time spent above half-maximum fluorescence intensity (double arrow). The values of each parameter are indicated for the specific curves shown (assembly rate constant in green, disassembly rate constant in red, and lifetime in black). (C) Quantification of assembly rate constants (left), disassembly rate constants (middle), and lifetime (right) for FAs in cells expressing shCTRL, shATG7, or shATG12. Data presented as median (line), first and third quartile (box), and whiskers extend to ±1.5 times the interquartile range. Individual data points outside of this range are shown. n = 64 FAs for shCTRL, n = 62 FAs for shATG7, and n = 51 FAs for shATG12, pooled from four independent experiments. P-values calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. n.s., not significant.