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. 2006 May 8;173(3):383–394. doi: 10.1083/jcb.200511093

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Copyright © 2006, The Rockefeller University Press

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Figure 5. — Mechanism of transverse arc assembly. (A) A U2OS cell expressing α-actinin–CFP (red) and YFP-MLC (green) was monitored by time-lapse imaging. Individual α-actinin (a) and myosin (m) bands are indicated by numbers (see diagram on the right). Cell edge is located on the left side and the center of the cell on the right side. Transverse arcs are generated by endwise annealing of short α-actinin cross-linked actin and myosin bundles. Bar, 5 μm. (B) U2OS cells were treated with 90% DMSO (control) for 30 min (top) or with 50 μM blebbistatin for 20 min (middle) or 30 min (bottom). Cells were fixed, and F-actin, vinculin (focal adhesions), and myosin II were visualized by phalloidin and anti-vinculin and anti–myosin II antibodies, respectively. White arrows indicate remaining focal adhesions and dorsal stress fibers in blebbistatin-treated cells, whereas transverse arcs were no longer detected in these cells after 20 min of blebbistatin treatment. Bars, 10 μm.