Journal: The Journal of Biological Chemistry

Article Title: S-nitrosylation of the scaffold protein STRAP enhances oxidative stress–induced apoptosis

doi: 10.1016/j.jbc.2026.111141

Figure Lengend Snippet: Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.

Article Snippet: Eluted proteins were separated by SDS-PAGE on 4% to 20% Criterion Precast Midi Protein Gels (Bio-Rad), transferred to PVDF membranes, and probed with the following antibodies: rabbit anti-eNOS (Cell Signaling, 32027S), rabbit anti-nNOS (Cell Signaling, 4231S), rabbit anti-iNOS (Santa Cruz, sc-8310), and rabbit anti-myc (Cell Signaling, 2278S).

Techniques: Expressing, Immunoprecipitation, Control, Western Blot, Negative Control