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. 2011 Feb 15;286(13):11179–11184. doi: 10.1074/jbc.M110.201780

FIGURE 3.

FIGURE 3.

Thioredoxin interacts with NSF in the presence of NO. A, interaction between endogenous NSF and endogenous TRX1. HUVEC were treated with SNAP (500 μm) or l-NAME (500 μm) for 4 h. Cell lysates were immunoprecipitated with antibody to NSF and immunoblotted for TRX1. NSF interacts with TRX1 in an NO-dependent manner. B, quantitation of A (mean ± S.D). **, p < 0.01. C, stable interaction between NSF and TRTX1 depends upon TRX1 residue Cys-35. HeLa cells were transfected with a vector expressing wild-type Myc-TRX1(WT) or mutant Myc-TRX1(C35S) and treated with SNAP for 4 h. Cell lysates were immunoprecipitated with antibody to Myc and immunoblotted for NSF. TRX1 interacts with NSF, through the CXXC disulfide reductase domain of TRX1. D, HeLa cells were transfected with a vector expressing mutant Myc-TRX1(C35S) and treated with increasing amounts of SNAP (5–500 μm) for 4 h. The interaction between NSF and TRX1 was analyzed as above. SNAP increases the interaction in a dose-dependent manner.