Figure 1. Impaired migration rate and increased FA size in autophagy-deficient cells. (A) Representative phase-contrast microscopy time-lapse sequences of single cells expressing shControl (CTRL; left), shATG7 (middle), or shATG12 (right) tracked over 3 h after wounding. Elapsed time (h) in top left of images. Bars, 10 µm. These images correspond to Video 1. (B) Migration paths of individual shCTRL (left), shATG7 (middle), or shATG12 (right) cells showing total distance traveled over 3 h. Cell position over time was used to generate paths and was determined by manually tracking cell nucleoli in each frame over the course of the time lapse. n = 10 representative cells shown per condition, and each colored track represents an independent cell. The starting position for each cell was normalized to 0 µm, 0 µm on the x, y axes. (C) Quantification of migration rate of individual tracked cells determined as total distance traveled divided by the total time of migration (d/tf – t0). 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 = 155 cells for shCTRL, n = 121 cells for shATG7, and n = 115 cells for shATG12, pooled from three independent experiments.P-values calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. (D) Representative immunofluorescence images of migrating wound edge cells expressing shCTRL (top), shATG7 (middle), or shATG12 (bottom) stained for endogenous F-actin (green) and paxillin (magenta) to mark FAs. Right panels show enlarged insets of boxed region in merged images. Bars, 5 µm. Insets are magnified 3.7-fold. (E) Quantification of the area of leading edge FAs in migrating wound edge cells determined by manually outlining anti-paxillin-labeled FAs. Data presented as median (line), first and third quartile (box), and whiskers extend to ±1.5 times the interquartile range. n = 713 FAs for shCTRL, n = 511 FAs for shATG7, and n = 430 FAs for shATG12, pooled from two independent experiments. P-values calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. n.s., not significant.
Figure 1.

Impaired migration rate and increased FA size in autophagy-deficient cells. (A) Representative phase-contrast microscopy time-lapse sequences of single cells expressing shControl (CTRL; left), shATG7 (middle), or shATG12 (right) tracked over 3 h after wounding. Elapsed time (h) in top left of images. Bars, 10 µm. These images correspond to Video 1. (B) Migration paths of individual shCTRL (left), shATG7 (middle), or shATG12 (right) cells showing total distance traveled over 3 h. Cell position over time was used to generate paths and was determined by manually tracking cell nucleoli in each frame over the course of the time lapse. n = 10 representative cells shown per condition, and each colored track represents an independent cell. The starting position for each cell was normalized to 0 µm, 0 µm on the x, y axes. (C) Quantification of migration rate of individual tracked cells determined as total distance traveled divided by the total time of migration (d/tf – t0). 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 = 155 cells for shCTRL, n = 121 cells for shATG7, and n = 115 cells for shATG12, pooled from three independent experiments.P-values calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. (D) Representative immunofluorescence images of migrating wound edge cells expressing shCTRL (top), shATG7 (middle), or shATG12 (bottom) stained for endogenous F-actin (green) and paxillin (magenta) to mark FAs. Right panels show enlarged insets of boxed region in merged images. Bars, 5 µm. Insets are magnified 3.7-fold. (E) Quantification of the area of leading edge FAs in migrating wound edge cells determined by manually outlining anti-paxillin-labeled FAs. Data presented as median (line), first and third quartile (box), and whiskers extend to ±1.5 times the interquartile range. n = 713 FAs for shCTRL, n = 511 FAs for shATG7, and n = 430 FAs for shATG12, pooled from two independent experiments. P-values calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. n.s., not significant.

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