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. 2000 May 29;149(5):1097–1106. doi: 10.1083/jcb.149.5.1097

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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(A) Immunoblot of total cell proteins of WT, βDDDE440 (+ α-COOH terminus-β chimera) and βDDDD₄₄₀ (+ α-COOH terminus β) chimera mutants probed with the polyclonal anti-polyG antibodies (top), monoclonal anti–β-tubulin antibodies, DM1B (middle), and monoclonal anti–α-tubulin antibodies, DM1A (bottom). Approximately equal amounts of total cell proteins were loaded in each lane. (B) A Western blot of ciliary proteins of the βDDDDGD₄₄₂ (+ α-COOH terminus-β chimera) and βDDDE₄₄₀ (+ α-COOH terminus-β chimera) mutants probed with several antitubulin antibodies. Equal amounts of total proteins were loaded into each lane. The same blot was probed sequentially using the ECL system. After each exposure, the antibodies were removed, and the blot was reprobed according to manufacturer's instructions. The following antibodies were used: anti–β-tubulin, DM1B; anti–α-tubulin, DM1A; antimonoglycylated tubulin, TAP 952; and antipolyglycylated tubulin, AXO 49.