Fig. 5

RhoH depletion alters migration behaviour of PC3 cells. RhoH depletion reduces cell migration speed. PC3 cells were transfected with the indicated siRNAs targeting RhoH or control siRNA. Cells were seeded in 24-well dishes 48 h after transfection and monitored by time-lapse microscopy for 24 h. a Example cell tracks are shown for each condition, plotted with their starting point at the intersection of the x- and y-axes. b Quantification of PC3 cell migration speed. At least 50 cells were tracked from three independent experiments. Boxes of box and whisker plots show median, 25th and 75th percentiles; whiskers show minimum and maximum values; ***p ≤ 0.001, Student’s t test. c Effect of RhoH on HGF-induced migration. 48 h after siRNA transfection, cells were transferred to 0.1% FCS with or without 20 ng/ml HGF. Cells were monitored by time-lapse microscopy for 24 h, with an image taken every 4 min. At least 100 cells were tracked from three independent experiments. Boxes of box and whisker plots show median, 25th and 75th percentiles; whiskers show minimum and maximum values; **p ≤ 0.01, ***p ≤ 0.001, Student’s t test. d Data for State 1 and State 2, showing mean migration speed, directional variance (over a time period of 8 min) and proportion of total time spent in State 1 across all cells (α). e Analysis of cell tracks indicated two modes of migration, State 1 and State 2, with different migration speeds and persistence (directional variance)