Please also see supplemental Figure S5C.
Immunoblotting analyses were performed with the indicated antibodies.
(A, B) WT rH3 or rH3-T11A expression was reconstituted in endogenous H3-depleted U87/EGFR cells (A), which were then treated with or without EGF (100 ng/ml) for 6 h. ChIP analyses with a HDAC3 antibody were performed (B).
(C) GST-HDAC3 pull-down assay was performed by incubation of 100 ng of purified recombinant His-tagged WT histone H3 or H3-T11A mutant with or without immobilized GST-HDAC3, which was followed by incubation with 200 ng of purified recombinant WT His-PKM2 or His-PKM2 K367M in the presence of PEP.