Fig. 4
Effect of chemical perturbations on wound response. a Region of interest and 1-D coordinate system defined along the membranellar band. Scale bar 20 μm. b Kymographs and c 2-D autocorrelations of the kymographs for cells under control (no treatment), NiCl2, or KI treatment, before and after wounding in Regime 2. Analysis of representative cells shown (N = 6 per experimental group). d Measured timescales of mechanical modes of wound response in untreated control (PSW), NiCl2-treated, and KI-treated cells, shown as box and whisker plots with a line at the median. Data in each treatment condition combined from Regimes 1 and 2; individual cells shown as dots (control: N = 16–21 cells per mode; NiCl2: N = 6–22 cells per mode; KI: N = 8–19 cells per mode). Timescales were measured as described in the “Imaging of cell motion after wounding” section. Significance values were obtained from multiple t tests for treated cells compared to untreated cells, α = 0.05. Only the NiCl2-treated twisting/pulling timescale was significantly different from that of untreated cells (p = 0.0006). All other timescales were not significantly different (NS, p > 0.05). e Relative occurrences of mechanical modes of wound response across wounding regime and treatment condition (Total N = 25–31 cells per experimental group). f Fraction of NiCl2-treated cells healed, using the wound repair assay. Data shown as mean (stars) of N ≥ 3 biological replicates (dots) and fit to a one-phase exponential function using mean datapoints in addition to the 24-h (86,400 s) survival rate, to aid extrapolation. Untreated (PSW) curves included for comparison. The control line shows the mean fraction of unwounded NiCl2-treated control cells below Ithreshold, and the shaded region indicates SD (3 biological replicates). Total N = 26–59 cells per experimental condition. Survival rate of g NiCl2-treated cells and h KI-treated cells. Data for unwounded control cells, 24 h after washing, and Regime 1 or 2 wounded cells, 24 h after wounding, shown as mean (bar) of 3 biological replicates (dots) (N = 15–47 cells per replicate)