Autophagy inhibition results in enhanced cell spreading. (A) Spinning disk confocal microscopy time-lapse sequences of cells expressing ZsGreen during spreading after replating. Representative shCTRL (top), shATG7 (middle), or shATG12 (bottom) cells shown over a 3-h time course. Elapsed time (h) indicated in top left of images. Bars, 10 µm. (B) Representative images of ZsGreen-expressing cells fixed 1 h after replating used for quantification of cell area in C. Whole field images shown with enlarged boxed insets of individual cells at bottom left. Tracing of individual cell in inset shown at bottom right. Bars, 50 µm. Insets are magnified 2.4-fold. (C) Quantification of area of cells fixed 1 h after replating. Area determined by manually outlining individual ZsGreen-expressing cell borders. 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 = 315 cells for shCTRL, n = 351 cells for shATG7, and n = 306 cells for shATG12, pooled from three independent experiments. P-values were calculated using a nonparametric Kruskall-Wallis test followed by Dunn post-hoc test. n.s., not significant.