Supplementary Fig. S3.

S-nitrosylation of the ASK1 recombinant protein by NO donors, GSNO and DEANO. ASK1 recombinant protein was incubated with increasing concentrations of (A) GSNO and (B) DEANO for 15 min and subjected to fluorescence switch or biotin switch assay, respectively. S-nitrosylated proteins were detected by their fluorescent maleimide signal (A, upper panel) or immunoblot using an anti-biotin antibody (B, upper panel). Coomassie and Sypro Ruby staining are shown as a loading control (lower panels) in A and B, respectively. The experiment was repeated at least three times with similar results.