Figure 5.
Effect of redox state on the thioredoxin-Txnip interaction. (A) Thioredoxin-C73S-GST beads were incubated in 100 μM of dithiothreitol (DTT) or hydrogen peroxide (H2O2) for two hours and washed 3 times in at least 20 bed volumes of lysis buffer. The beads were then incubated with cell lysates containing wildtype Txnip or C247S Txnip as a negative control. Bound proteins were probed for Txnip with JY2. (B) Lysates containing wildtype or C247S Txnip were added to unmodified thioredoxin-C73S-GST beads in the presence or absence of 100 μM DTT and H2O2. Bound proteins were probed for Txnip. The experiments were replicated with essentially identical results as those shown.