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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Free Radic Biol Med. 2021 Feb 17;166:73–89. doi: 10.1016/j.freeradbiomed.2021.02.006

Figure 9.

Figure 9.

MPO Oxidation of KYC Increases KYC Thiylation of Extracellular HMGB1. (A) Streptavidin affinity-blot of conditioned media from RLMVEC cultures treated with media, media + B-KYC, and media + MPO reaction system (MPO = 120 nM; H2O2 = 50 μM) + B-KYC. The affinity-blot shows that the MPO reaction system + B-KYC predominately thiylates HMGB1 released into the conditioned media relative to the immunoblot levels of HMGB1 protein released (immunoblot for HMGB1). (B) Streptavidin affinity-blot of cell lysates of RLMVEC cultures treated with media, media + B-KYC, and media + MPO reaction system (MPO = 120 nM; H2O2 = 50 μM) + B-KYC. This affinity-blot shows that the MPO reaction system + B-KYC thiylates low levels of HMGB1 in cell lysates. This conclusion is based on the relative levels of KYC thiylated HMGB1 in panel A vs. KYC thiylated HMGB1 as a function of HMGB1 protein in conditioned media vs. RLMVEC cell lysates.