Fig. 2

ROCK inhibition significantly decreases BMSC polarisation and migration. a ROCK inhibition (10 μM Y27632 pre-treatment for 3 h) significantly reduces serum-starved BMSC polarisation in response to an MCP-1 gradient in a 2D model. Permeabilised serum-starved BMSCs immunofluorescently stained with F-actin (red) and α-tubulin (green) and DNA stained with DAPI (blue), following the indicated treatments. Scale bar = 10 μm. b, c ROCK inhibition (10 μM Y27632 combined with SFM for 24 h) significantly reduces MCP-1-stimulated BMSC migration. b ROCK inhibition (10 μM Y27632 combined with SFM for 24 h) significantly reduces BMSC migration in a three-dimensional assay in response to MCP-1. Graph represents three separate independent experiments. Data are presented as mean ± standard deviation. The migration index (MI) is a relative change in measured electrical impedance. When there are no cells, MI is 0. Increasing MI is a quantitative measure of cells attached to the electrodes, representative of cell numbers migrating though a membrane. c, d ROCK inhibition (10 μM Y27632 combined with SFM for 24 h) significantly reduces BMSC 2D migration in mouse (c) but not human (d) cells in response to 3-h MCP-1 exposure. SFM set = 1. Graphs represent three independent experiments (>100 cells evaluated for each separate independent experiment). Data are presented as mean ± standard error of the mean. A P value of less than 0.05 (*) was deemed significant, and a P value of less than 0.001 (***) highly significant. 2D two-dimensional, BMSC bone marrow-derived stromal cell, DAPI 4',6-diamidino-2-phenylindole, MCP-1 monocyte chemoattractant protein 1, ROCK Rho-associated, coiled-coil containing protein kinase, SFM serum-free medium