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. 2017 Mar 31;292(22):9063–9074. doi: 10.1074/jbc.M116.771832

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 5. — Prereduced and preoxidized CGPC and CGRC form reduction-dependent complexes with substrates from activated washed platelets. Platelet releasates (A and B) and lysates (C and D) were prepared as described under “Experimental procedures.” Samples were then immunoprecipitated with anti-FLAG antibody, purposefully overloaded to detect PDI-substrate complexes, and separated by SDS-PAGE in either non-reducing (A and C) or reducing (B and D) conditions followed by immunoblotting using anti-FLAG antibody. The catalytically inactive AGHA variant was used as a control. Lane 1, prereduced CGPC-PDI; lane 2, prereduced CGRC-PDI; lane 3, prereduced AGHA-PDI; lane 4, preoxidized CGPC-PDI; lane 5, preoxidized CGRC-PDI; lane 6, preoxidized AGHA-PDI. Molecular mass standards are shown on the left and right.