Migration of MTLn3 carcinoma cells in HD matrix. Tumour cells were seeded on matrices for 24 h and treated with vehicle, the rock inhibitor (RI), Y-27632, and the MMP inhibitor, GM6001, and allowed to migrate for a further 48 h. The cells were stained with phalloidin-Alexa488 and imaged by confocal and reflection microscopies to capture cells and collagen, respectively. The image stacks were reconstructed in the x-z plane to measure the depth of cell migration. A, The RI blocked cell migration in 1 mg/cm3 matrix but had no significant effects on cell migration in higher density matrices of 10 and 20 mg/cm3. B, GM6001 significantly inhibited cells migration at higher concentrations but had no effect at the lowest concentration of 12.5 μm. C, Cell migration was significantly inhibited in the presence of both RI (10 μm) and low concentrations of GM6001 (12.5 μm) compared to vehicle-treated cells alone. Grey and white boxes show the upper and lower quartiles, respectively. Upper bars represent the maximum and lower bars, the minimum distance travelled by cells. Each experiment was repeated at least twice. 1-way ANOVA followed by post-hoc Tukey’s test indicating significant difference at * p < 0.05 and ** p < 0.01.