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Figure 4. PDS5 is found in association with SA1- and SA2-containing cohesin complexes. (A) Low-speed supernatant of extracts from logarithmically growing HeLa cells was analyzed by immunoprecipitation with either preimmune (P) or immune antibodies (I) to PDS5. Immunoprecipitates (IP) and supernatants (SUP) were analyzed by SDS-PAGE and Western blotting with antibodies to the indicated proteins. (B) HeLa extracts prepared as in A were analyzed by immunoprecipitation with SA1, SA2, or with nonspecific control (MOCK) antibodies and the immunoprecipitates (IP), extracts before immunoprecipitation (Input) and supernatants (Sup) were analyzed by SDS-PAGE and Western blotting with antibodies to PDS5. (C) HeLa extracts prepared as in A were analyzed by immunoprecipitation with either preimmune (P) or immune (I) SA2 antibodies (446). After washing with buffers containing either 150, 250, or 500 mM NaCl, the immunoprecipitates were analyzed by SDS-PAGE and Western blotting with antibodies to the indicated proteins. (D) Xenopus interphase extracts were analyzed by immunoprecipitation with either nonspecific (IP-contr), Xenopus PDS5 (IP-PDS5), or SA1 (IP-SA1) antibodies. The precipitates were analyzed by immunoblotting with PDS5, SA1, and SCC1 antibodies. After immunoprecipitation with control (xtΔcontr), PDS5 (xtΔPDS5), or SA1 (xtΔSA1) antibodies the resulting supernatants were analyzed side by side. PDS5 antibody 648 was used for the IP and 647 for immunoblotting. Image published in: Sumara I et al. (2000) © 2000 The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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