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. 2010 Aug 9;190(3):331–345. doi: 10.1083/jcb.201003005

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© 2010 Ong et al.

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Figure 1. — p125A binds to Sec31A. (A) GST-Sec31A and GST immobilized onto glutathione beads were incubated with rat liver cytosol and proteins retained on the beads were eluted and resolved on a 7% SDS-PAGE gel. Distinct bands were cut out and subjected to protein identification by mass spectrometry. Arrow indicates mammalian p125A with the identified peptide sequences shown. (B) myc-p125A and GFP-Sec31A were exogenously expressed in HEK293 cells either singly or in combination as indicated. Cell lysates were prepared and subjected to immunoprecipitation by either anti-myc (9E10) antibodies or control mouse IgG. The samples (together with 5% starting materials as controls) were resolved by SDS-PAGE and transferred to PVDF and immunoblotted using antibodies against GFP and myc. Molecular size markers are in kD.