The peptide pull-down assays were performed in accordance with the protocol described67 (link). In brief, 1.0 µg of biotinylated histone peptides were incubated with 1 μg of protein or 0.5 ml whole cell extract in binding buffer (50 mM Tris-HCl, pH 7.5, 250 mM NaCl, 0.1% NP-40, 1 mM PMSF, protease inhibitors) overnight at 4 °C. After 1 h incubation with Streptavidin beads (Amersham), the beads were washed with the binding buffer and boiled for 5 min. The supernatant from boiled beads was subject to immunoblot analysis. The peptides used are the following:
Unmodified H3 (1–23): ARTKQTARKSTGGKAPRKQIASK;
H3K4me3 (1–23): ARTK(me3)QTARKSTGGKAPRKQIASK;
H3pT11 (1–23): ARTKQTARKST(p)GGKAPRKQIASK;
Unmodified H3 (72–85): REIAQDFKTDLRFQ;
H3K79me3 (72–85): REIAQDFK(me3)TDLRFQ.
Unmodified H3 (1–23): ARTKQTARKSTGGKAPRKQIASK;
H3K4me3 (1–23): ARTK(me3)QTARKSTGGKAPRKQIASK;
H3pT11 (1–23): ARTKQTARKST(p)GGKAPRKQIASK;
Unmodified H3 (72–85): REIAQDFKTDLRFQ;
H3K79me3 (72–85): REIAQDFK(me3)TDLRFQ.
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