Figure 7.

Effects of Oxidative Stress on KYC Thiylation of RLMVEC Proteins. (A) Streptavidin affinity-blot of KYC thiylated proteins from cell lysates prepared from RLMVEC cultures treated with B-KYC (baseline), B-KYC+MPO+H2O2 (MPO-dependent), and B-KYC+H2O2 (H2O2-dependent). (B) Immunoblot for Nrf2 in RLMVEC cultures treated as described in A. (C) Immunoblot for Keap1 in RLMVEC cultures treated as described in A. (D) Immunoblot for HMGB1 in RLMVEC cultures treated as described in A. (E) Immunoblot for β-Actin in RLMVEC cultures treated as described in A. (F) Bar chart showing relative levels of KYC thiylation, Nrf2, Keap1, and HMGB1 as a function of β-Actin (n=2, * = p<0.05, statistical analysis by ANOVA) These data show that MPO-dependent oxidation of KYC increases KYC thiylation in RLMVEC proteins cells that are proximal to MPO, while H2O2-dependent oxidation induces a slight, if any, increase in KYC thiylation of RLMVEC proteins.