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. 2013 Jan 9;288(9):6053–6062. doi: 10.1074/jbc.M112.433284

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — JMJD6 interacts with and hydroxylates histone H3 and H4 in vitro. A and B, in vitro pulldown assay. Biotin-labeled histone H3_1–21 peptides (A) or recombinant histone H4 (B) were incubated with or without GST-JMJD6, pulled down by streptavidin-Sepharose, and detected by dot blot using streptavidin-HRP (A) or Coomassie Brilliant Blue (CBB) staining (B). Pulled down GST-JMJD6 was detected by Western blotting using anti-JMJD6 antibody (A) or Coomassie Brilliant Blue staining (B). C and D, enzymatic activity of GST-JMJD6 was measured by MS analysis. Histone H3_1–20 (C) and H4_1–30 (D) peptides were served as substrates. BSA was used as a negative control. IB, immunoblot.