Fig. 7. Microtubuli repolymerization assays in mutant and wild-type RII
-transfected cells. Reh, Reh-RII and Reh-RII(T54E) cells were incubated for 1 hour at 4°C to depolymerize microtubules. Repolymerization was started by heating to 37°C and samples were stopped at different time points (0-15 minutes) by methanol fixation. Aster formation was then analyzed by immunofluorescence using anti--tubulin mAb. Mitotic (A) and interphase (C) cells were examined. The right-hand columns show staining by anti-RIIß antibody in cells prior to depolymerization. Arrows indicate the centrosomal association of RIIß. (B) The number of mitotic cells that formed asters after depolymerization was scored. Reh vs RII, P<0.005; Reh vs RII(T54E), P<0.001.
