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. Author manuscript; available in PMC: 2017 Jul 19.
Published in final edited form as: Nat Mater. 2016 Mar 14;15(7):792–801. doi: 10.1038/nmat4586

Figure 5.

Figure 5

Modulation of topotaxis direction of SBcl2 cells by perturbations of PI3K and ROCK signaling. (a) Predicted changes of activity of the signaling network regulating cell stiffness of SBcl2 cells by pharmacological inhibition of PI3K (LY294002, red arrow) and ROCK (Y27632, blue arrow). PI3K inhibition is predicted to extend the reversal of the balance between PI3K and ROCK activities to the whole ECM density range in contrast to ROCK inhibition removing the reversal; (b,e) SBcl2 cell displacements in the direction parallel to post density gradient on 10 μg/ml FN pre-coated GPDA (b) and 50 μg/ml FN pre-coated GPDA examined for 5 hours (e). * = Statistical significance of the biased deviation from the zero mean in the x-direction. **P<0.01 and ***P<0.005. Wilcoxon signed rank test for the zero mean. All error bars are S.E.M.; (c,f) The distributions of topotactic SBcl2 cell displacements on 10 μg/ml FN pre-coated GPDA (c) and 50 μg/ml FN pre-coated GPDA (f) following treatment with a ROCK inhibitor, (5μM of Y27632) and a PI3K inhibitor (10μM LY294002); (d,g) Modulation of cell stiffness dependence on the post density of 10 μg/ml FN (d) and 50 μg/ml FN coated GPDA (g) following PI3K and ROCK signaling perturbations. * = Statistical significance between the values on the dense vs. sparse regions. *P<0.05 and all paired two-sample Student's t-test. All error bars are S.E.M. (h) Modulation of topotaxis direction of non-invasive melanoma cells cultured on GPDA pre-coated with a high FN density, following inhibition of PTEN. SBcl2 and WM1552 cell displacements in the direction parallel to post density gradient on GPDA pre-coated with 50 μg/ml FN and examined for 5 hours following treatment with 100 nM of bpV(HOpic), a PTEN inhibitor. * = Statistical significance of difference in displacement in the x-direction between with and without drug. ***P<0.005. All paired two-sample Student's t-tests. All error bars are S.E.M. (i) The distributions of topotactic SBcl2 and WM1552 cell displacements measured in (h). * = Statistical significance of the biased deviation from the zero mean in the x-direction. ***P<0.005. Wilcoxon signed rank test for the zero mean.