File:Epithelial cell microfilaments 01.jpg
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Organization and dynamics of F-actin in migrating newt lung epithelial cells
(a and b) Z-series of deconvolved optical sections of an Alexa-488 phalloidin-stained cell were used to generate views along the z axis (a, maximum projection) or y axis (b, three-dimensional reconstruction of the boxed area in panel a). Bar, 10 μm.
(c and d) Confocal FSM image, taken from a time-lapse series, of a living cell injected with X-rhodamine actin (c) and after fixation and staining with Alexa 488 phalloidin (d). See Video 1 (available at http://www.jcb.org/cgi/content/full/jcb.200203022/DC1). F-actin bundles (c and d, arrowheads) prominent in the phalloidin image (c) correspond to linear arrays of speckles in the FSM image (d, boxed bundle shown at 150-s intervals in inset). A bundle in d (starred arrowheads) has no corresponding linear array of speckles in the FSM image (c). Bar, 10 μm; inset, 3 μm.
(e–i) Four zones of f-actin dynamics. (e) Single f-actin FSM image from Video 2 (available at http://www.jcb.org/cgi/content/full/jcb.200203022/DC1). Bar, 10 μm. (f–i) Kymographs of the four zones of f-actin movement taken along the regions highlighted in (e). F-actin speckles move retrograde in the (f) lamellipodium at 1.6 μm/min, in the (g) lamella at 0.3 μm/min. (h) In the convergence zone, there is no directional movement of f-actin. (i) F-actin moves anterograde in the cell body at 0.4 μ/min. Elapsed time in min:s. Bar, 10 μm.
(j) Plot of the actin:fluorescent volume marker ratio along the axis of migration of a cell reveals a steep gradient of actin concentration across the lamellipodium (lp), and relatively constant concentration through the rest of the cell.
Original file name: Figure 1. http://jcb.rupress.org/content/158/1/31/F1.large.jpg
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|current||09:59, 28 March 2011||913 × 973 (289 KB)||S8600021||==Organization and dynamics of F-actin in migrating newt lung epithelial cells== (a and b) Z-series of deconvolved optical sections of an Alexa-488 phalloidin-stained cell were used to generate views along the z axis (a, maximum projection) or y axis (b,|