Fig. 5: S-nitrosylation of the CAT protein in vitro and in vivo.

a Recombinant CAT was treated with GSH (negative control for GSNO) or GSNO to measure S-nitrosylation by the biotin switch method. Without sodium ascorbate (Asc) sample as another negative control. Protein samples were separated via SDS–PAGE and analyzed by immunoblotting using an anti-biotin antibody and the His tag antibody. b Protein extracts were prepared from WT and GSNOR-silenced strains, and the proteins were analyzed by the biotin-switch assay. The protein was incubated with streptavidin beads, which were then washed to detect the S-nitrosylation of the CAT protein in the WT and GSNOR-silenced strains. Equal loading was verified by antibodies specific for actin and CAT.