Fig. 8. Visualization of ß-tubulin in lens epithelial wholemounts by immunofluorescence and confocal microscopy in different cell-cycle phases. Seven-day-old wild-type or A^–/– mouse lens epithelial wholemounts were immunostained with an antibody to ß-tubulin (red) and the nuclei were stained with TOTO-1 (green). (A) In the interphase of wild-type cells, ß-tubulin was uniformly distributed around the nuclei. (B,C) In metaphase cells, the bundling of ß-tubulin around the condensed chromosomes could be readily detected. Note that there was no difference in the organization of the metaphase spindles of the wild-type (B) and A^–/– (C) lens epithelial wholemounts. (D-F) In anaphase cells of the wild-type epithelium, the spindle was well organized (D). However, there was disorganization of the anaphase spindle of the A^–/– lens epithelial wholemounts (E,F). This aberrant spindle phenotype was observed in 45% of the anaphase spindles of the A^–/– wholemounts. Twenty metaphase and 20 anaphase spindles in six different lens epithelial wholemounts were analyzed for each genotype. Bars, 5 µM.