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Fig. 4. Syne-1 localizes to central spindle and midbody, but not to the contractile ring. Mitotic NRK cells, double-stained with anti-Syne-1 (red) (a,b,d,e,g,h) and either with anti-tubulin (green) (a-c,e) or with fluorescein phalloidin (green) (f,h). (a) Conventional fluorescence microscopy reveals that Syne-1 co-localizes with spindle-microtubules (arrows) and that its distribution during anaphase is restricted along the spindle to the central region. (b) High-resolution image generated by optical sectioning microscopy of an anaphase mitotic spindle, double stained with antibodies to tubulin (green) and Syne-1 (red), demonstrates that Syne-1 associates with the central spindle microtubules in the form of discrete puncta (c-e, arrows). An optical sectioning image of NRK cells entering cytokinesis double stained with antibodies to tubulin (c) and Syne-1 (d) (e is an overlay of c and d) illustrates that Syne-1 localizes to a ring-like structure that surrounds the spindle midbody (d and e, arrows). (f-h) Fluorescein phalloidin stained actin of the contractile ring (f, arrowhead), which encircled the Syne-1 positive structure at the midbody (g, arrow). An Overlay (h) of f and g revealed that Syne-1 (h, arrow) did not localize to the contractile ring (h, arrowhead).
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