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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 11.

Figure 11.

Effects of KYC on HMGB1 Thiol Oxidation State and Lysyl-Acetylation. (A) Representative immunoblots for cysteine sulfonyl on HMGB1 in lung lysates from normoxic and hyperoxic pups. Bar chart presents mean ± SD of relative levels of cysteine sulfonyl on HMGB1 in lung lysates from normoxic and hyperoxic pups. These data show that hyperoxia decreased the levels of cysteine sulfonyl on HMGB1 in lung lysates. (B). Representative immunoblots for cysteine sulfonyl on HMGB1 in lung lysates from hyperoxic pups treated with PBS or KYC. The bar chart presents mean ± SD of relative levels of cysteine sulfonyl on HMGB1 in lung lysates from hyperoxic pups treated with PBS or KYC. These data show that KYC treatment increased the levels of cysteine sulfonyl on HMGB1 in lung lysates of hyperoxic pups. (C) These data suggest that activated immune cells are the predominant source of lysyl-acetylated HMGB1. Dead and dying cells are the predominant source of non-acetylated HMGB1. Representative immunoblots for lysyl-acetylated residues on HMGB1 immunoprecipitated from lung lysates from hyperoxic pups. HMGB1 was immunoprecipitated with non-depleting concentrations of anti-HMGB1 antibody. The immunoblots show difference in lysyl-acetylated HMGB1 in lung lysates from hyperoxic pups with PBS and KYC treatments. The hyperoxic pups treated with PBS have predominantly non-acetylated HMGB1, which is released by dead and dying cells. In contrast, the predominant HMGB1 isoform in lung lysates from KYC treated hyperoxic pups is lysyl-acetylated HMGB1. These data demonstrate that HMGB1 release in lungs of hyperoxic pups is shifted from dead and dying lung cells in PBS-treated hyperoxic pups to activated immune cells in KYC-treated hyperoxic pups.