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Fig. 5. Cosedimentation of ODF2 with microtubules. (A,B) Microtubules were isolated from cos-7 cells expressing one out of four ODF2-GFP fusion proteins under conditions that promote microtubule polymerization. Proteins were separated using denaturing SDS-PAGE and transferred to Hybond-C. Probing with antibodies against GFP (A) revealed cosedimentation of the full-length ODF2NC as well as the truncated ODF2 proteins ODF2NC1, ODF2NC2 and ODF2N2C with microtubules. (B) Detection of tubulin by anti-{alpha}-tubulin antibodies in the same fractions as in A. (C) Control performed with cos-7 cells expressing ODF2NC-GFP demonstrating the dependence of ODF2 sedimentation on microtubules. No ODF2 proteins were found in the final pellet if the experiment was performed under microtubule destabilising conditions (2), whereas the cellular debris after the first centrifugation step do contain ODF2-GFP (1). Incubation with antibodies against GFP.