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Fig. 7. KIF3B tail domain blocks Syne-1 localization to the midbody. (a-d) NRK cells transfected with myc-tagged KIF3B tail domain (c,d) or with full-length myc-KIF3B as a control (a,b). Cells were fixed and stained with anti-myc (a,c) to identify transfected cells, and with polyclonal SN120 to localize Syne-1 (b,d). While Syne-1 was concentrated at the midbody in cells transfected with the control full-length KIF3B (arrow) (b), it was significantly depleted from the midbody in cells transfected with KIF3B tail (arrow) (d). (e) To quantify this result, NRK cells transfected with KIF3B tail (tail) or with transfected with full-length KIF3B (full-length) were subjected to digital image analysis (see Materials and Methods). The amount of Syne-1 staining at the midbody (mean pixel density) was measured and is plotted as the ratio of midbody localized Syne-1 staining to background Syne-1 staining. Staining was measured in the cytoplasm directly adjacent to the midbody (error bars=standard deviation; * indicates the statistically significant difference from mock transfected cells by Student's t-test).
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