Figure 8.

MT affinity of XMAP4 and mutants. Interphase (I) and mitotic (M) cell extracts of nontransfected A6 cells (−) and A6 cells expressing GFP–WT, GFP–CM10A, GFP–C6A, or GFP–CM8A were incubated with Taxol-stabilized MTs and then centrifuged. XMAP4 coprecipitated with MTs (P) or remaining in the supernatant (S) was detected by immunoblotting with anti-XMAP4 antibody. The arrowhead and arrow denote endogenous XMAP4 and GFP–XMAP4, respectively. Approximately 70% of GFP–WT remained in the supernatant in mitosis, whereas ∼90% of GFP–CM10A, GFP–C6A, and GFP–CM8A coprecipitated with MTs.