Effects of RhoA GEFs depletion on intercellular communication. (A–C) Kymographs of speed and directionality for control versus ARHGEF18, ARHGEF3, and ARHGEF11 knockdown. (D) Vector fields 100 min after wound infliction for control versus ARHGEF11 knockdown. Bars, 100 µm. (E) Kymographs of speed and directionality for control versus ARHGEF28 knockdown. (F) Difference in PC3 for speed (top) and directionality (bottom) between GEF-H1 knockdown (two hairpins with 30% and 45% depletion) and control. Each point represents the difference in one well location and the mean of the same day’s control experiment. N = 1 d, n = 8 locations. (G) Validation of screen hits in RhoA GEFs. Each point represents the difference in PC of the indicated variable between GEF knockdown in one well location and the mean of the daily control; N, number of wells; n, number of locations. RHOA: N = 9, n = 47; ARHGEF18: N = 6, n = 31; ARHGEF11: N = 6, n = 23; ARHGEF28: N = 10, n = 53; ARHGEF3: N = 9, n = 48. Statistics via Wilcoxon signed rank test: *, P ≤ 0.01; **, P ≤ 0.001; ***, P ≤ 0.0001.