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. 2006 Jan 16;172(2):259–268. doi: 10.1083/jcb.200506179

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Copyright © 2006, The Rockefeller University Press

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Figure 3. — Stress fiber–derived α-SMA accumulates in detergent-soluble rods. REF-52 myofibroblasts were grown for 12 h on 4- (A) and 20-μm-long (B and D) adhesion islets and stained for α-SMA (green), β-cytoplasmic actin (red), and FN (blue). By using a detergent-free staining procedure, α-SMA was identified in cytosolic rods after growth on 4-μm-long islets (A) and in stress fibers on 20-μm-long islets of control cells (B). Inhibition of Rho-associated kinase on 20-μm-long islets leads to the neoformation of α-SMA rods (D; and Video 1). Bar, 20 μm. (C) Western blotting of the TX-100–insoluble cytoskeleton demonstrates the decrease of specific contractile proteins in the TX-100–insoluble fraction with decreasing adhesion islet size (Table I). Representative results of one out of four independent experiments are presented. MLCK, myosin light chain kinase; NM, nonmuscle; SM, smooth muscle. Video 1 is available at http://www.jcb.org/cgi/content/full/jcb.200506179/DC1.