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. 2008 Jan 2;28(6):2023–2034. doi: 10.1128/MCB.02130-07

FIG. 2.

FIG. 2.

HMTase activity of RE-IIBP. (A) Core histones were used as substrates in the HMTase assay with GST-RE-IIBP and GST-RE-IIBP point mutants (C483A and R477A). Methylation levels were quantified via filter binding assay, and data are represented as raw counts per minute incorporated. (B) Purified GST-RE-IIBP, GST-RE-IIBP point mutants, and GST proteins were incubated with HeLa cell extracts and subjected to HMTase assays. GST-bound beads were used as the negative control. The lower panel represents GST-RE-IIBP pulled down and immunoblotted with anti-G9a and anti-EZH2 antibodies. HeLa cell extracts were used as the positive control. (C) A similar HMTase assay as that described in panel A was performed with RE-IIBP deletion mutants (GST-RE-IIBP-1 and GST-RE-IIBP-2). A schematic representation of the domain structure of the recombinant RE-IIBP deletion mutants is shown. Conc, concentration; WT, wild type.