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Changes in acetyl-CoA levels and in histone acetylation are detected after manipulation of the mitochondrial pool of acetyl-CoA in 143B cells. Changes in acetyl-CoA levels and in histone acetylation are detected after manipulation of the mitochondrial pool of acetyl-CoA in 143B cells. (A) 143B rho+ cells were exposed for 4 h to BTC, a mitochondrial citrate carrier inhibitor. Total levels of acetyl-CoA were then estimated using deproteinized samples with a fluorescence-based assay and an acetyl-CoA standard curve; N = 3. t test was used to define statistical differences; error bars represent ±SD. (B) Histones were extracted and Western blots probing different histone acetylation marks performed after treatment of rho0 cells for 4 h with DM-α-KG or DCA; Westerns blots are representative. (C) Representative Western blots probing different acetylated histone residues upon treatment of rho+ or rho0 cells with HC, an inhibitor of the cytosolic ACL. Oswaldo A Lozoya et al. LSA 2019;2:e © 2019 Lozoya et al.
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