Fig. 7.

Suppression of Rho signalling through ROCK-inhibition leads to an increase in cell migration but not intrinsic cell directionality. Cell speed and directionality were calculated from time lapse images acquired with transmitted light microscopy before treatment and 1-hour post-treatment with 20 µM of the ROCK inhibitor or ddH₂O. (A) Average cell speed of CHO-K1 cells expressing paxillin-EGFP or co-expressing paxillin-EGFP and βPIX-mCherry before (blue) and after (red) treatment with Y-27632 or ddH₂O. Rose plots of migration tracks of CHO-K1 cells (B) expressing paxillin-EGFP, (C) co-expressing paxillin-EGFP and βPIX-mCherry before (top panels) and 1 h after (bottom panels) addition of Y-27632, or (D) control paxillin-EGFP expressing cells before (top panel) and after addition of ddH₂O (bottom panel) in culture media. For the Rose plots, each cell track starts at the 0,0 position. (E) Visual representation of how the directionality ratio was assessed from a cell’s migratory track, where d represents the net distance and D represents the total cumulative distance travelled by a cell along the entire track distance. (F) Cell directionality ratios of CHO-K1 cells expressing paxillin-EGFP or co-expressing paxillin-EGFP and βPIX-mCherry before and after treatment with Y-27632 or ddH₂O. Error bars are standard errors of the means. Four stars correspond to (****) p < 0.0001. n = 60–70 cells in total from 3 independent experiments. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)