Fig 7. Effect of disulfide reducing agents on the dissociation of Trx from Trx bound ASK1 and/or 14-3-3ζ complex.

The ASK1 signaling complex was co-IP using ASK1 or 14-3-3ζ antibody. Chemical dissociation of co-precipitated Trx from the complex was monitored in the presence or absence of various disulfide reducing agents in SDS- conditioned buffer as described in the text. DTT, dithiothreitol; 2-ME, 2-mercaptoethanol; THP, tris(hydroxypropyl)phosphine. The experiment was repeated at least twice and a representative Western blot is shown. A representative full blot image showing higher molecular weight protein complexes is shown in S1 Fig