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. 1999 Mar;10(3):713–726. doi: 10.1091/mbc.10.3.713

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Copyright © 1999, The American Society for Cell Biology

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S(all)A Clc1p associates with Chc1p in stable trimers. (A) Antibodies specific for Clc1p were used to coimmunoprecipitate Chc1p (Ip lanes) from extracts of S(all)A mutant cells (GPY1973, lane 1) or wild-type (WT) cells (GPY1970, lane 3). Total extracts (E lanes) representing ∼20% of the extract used for coimmunoprecipitation were analyzed directly in lanes 2 (GPY1973) and 4 (GPY1970). The band marked Ig corresponds to Ig heavy chain from the immunoprecipitates that reacts with the secondary antibody used in the immunoblot. (B) Sepharose CL4B column chromatography of extract from GPY1973 cells expressing S(all)A Clc1p. Extracts were prepared according to MATERIALS AND METHODS and fractionated by Sepharose CL4B column chromatography. Selected fractions were precipitated with 10% trichloroacetic acid and analyzed by immunoblotting with monoclonal antibodies to Chc1p or polyclonal antibodies to Clc1p. Column fraction numbers are indicated at the top of the panels. Wild-type clathrin displayed the same fractionation pattern.