Proliferation on soft matrices is decoupled from myosin light chain phosphorylation, cell tractions, and focal adhesions. Proliferation on soft matrices.

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Yvonne Aratyn-Schaus, Margaret L. Gardel Current Biology

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Volume 104, Issue 1, Pages (January 2013)

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Proliferation on soft matrices is decoupled from myosin light chain phosphorylation, cell tractions, and focal adhesions. Proliferation on soft matrices is decoupled from myosin light chain phosphorylation, cell tractions, and focal adhesions. (A) Immunofluorescence staining of phosphorylated myosin light chain in fibroblasts across matrix stiffness. Y27632 (10 µM) was applied for 24 hours. (B) Fibroblast root-mean-square tractions (RMST) measured by Fourier transform traction microscopy. Y27632 (10 µM) was applied for 45 minutes. Means and data points (n = 5 cells) are shown. Student's t-test: *P<0.0001 versus no drug. (C) Relationship between cell proliferation and tractions with or without Y27632 (10 µM). Proliferation is expressed as cell number relative to the no drug, rigid condition following 72 hours of culture of one representative experiment. (D,E) Immunofluorescence staining of vinculin (D) and talin (E) across matrix stiffness. Images are at 400× magnification; corresponding images of boxed regions are at 1000×. Scale bar: 25 µm. (F) Effect of matrix stiffness and Y27632 (10 µM) on focal adhesion (FA) length. Focal adhesions on 0.3 kPa gels were not observed above the detection threshold (0.2 µm). Data are means ± s.d. (n≥47). Student's t-test: *P<0.001 versus no drug. Justin D. Mih et al. J Cell Sci 2012;125:5974-5983 © 2012. Published by The Company of Biologists Ltd