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Fig. 3. The MTOC and stable MT subset do not reorient during EF motility, unlike wound-healing motility. (A) The total MT array (anti-ß-tubulin; a,c) and stable MT subset (detected via enrichment in detyrosinated tubulin; Glu, b,d) did not become polarized during EF-mediated motility (a,b), in contrast to wound-healing motility without applied EF (no EF; c,d), in which Glu MTs reoriented towards the wound edge. Glu MT labeling also allowed the detection of the MTOC (arrowheads, b,d). Migration direction is to the left in all micrographs; the cathode (-) (a,b) and wound edge (c,d) are shown with thin white arrows. Scale bar (d), 10 µm. (B) The proportion of cells with an oriented MTOC [i.e. within the cathode-facing quadrant (dotted lines)] was indistinguishable from the 25% expected for a random distribution. Similarly, 5.1% of cells in the EF showed a Glu MT array oriented towards the migration direction (i.e. with >75% of Glu MTs facing the migration direction), indistinguishable from controls (8.3%). Notice that, in the EF, only a small proportion of sparse cells had Glu MTs oriented in any direction. MTs are depicted as thin lines, Glu MTs as thick lines and the MTOC as a dot.
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