Fig. 3.

The native conformation of wild type HEDJ is similar to oxidized, diamide treated HEDJ and unlike reduced or Cys mutant HEDJ. A. Lysates of untransfected COS-1 cells (UT) were not treated (−) or were incubated with 5 mM diamide (O) or 20 mM DTT (R). Following incubation, 25 mM NEM was added to each sample to block sulfhydryl groups. Native gel sample buffer was added, samples were separated on a 7.5% Tris-glycine gel and immunoblotted using anti-HEDJ. B. Lysates from COS-1 cells that had been transfected with HEDJ-HA tagged constructs were treated and analyzed as in A. using mouse anti-HA antibody.