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Fig. 8. Silencing Cenp-F results in aberrant spindle morphology. (A) Transfected HeLa cells were fixed under conditions that preserve all microtubules and were then stained to detect tubulin and the DNA. Images show representative spindles in control and Cenp-F-deficient cells. Arrowhead identifies an abnormally long microtubule extending beyond the spindle midzone. (B) Histograms plotting tubulin intensity along the spindle axis. (C) Microtubule pelleting assay showing that whereas kinesin heavy chain (KHC) binds taxol-stabilized microtubules, Cenp-F does not. S, supernatant (i.e. the cell lysate used for the assay); C, the sucrose cushion; R, the ATP-release fraction; P, the microtubule pellet. The assay was performed with (+MTs) and without (–MTs) microtubules, confirming that the pelleting of KHC is microtubule dependent.





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