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. Author manuscript; available in PMC: 2014 May 16.
Published in final edited form as: Cell. 2013 Jul 18;154(2):297–310. doi: 10.1016/j.cell.2013.06.027

Figure 1. Human SET1C-Mediated H3K4 Trimethylation Is Stimulated by p53- and p300-Dependent H3 Acetylation on Recombinant Chromatin Templates.

Figure 1

(A) HAT and HMT assay scheme for p300 and SET1C.

(B) HAT assay with indicated factors and 3H-acetyl-CoA (autoradiographic analysis). Lower panels in (B) and (D) show loading controls stained with Coomassie brilliant blue (CBB).

(C) H3 site-specific HAT assays with indicated factors and acetyl-CoA (immunoblot analysis with antibodies indicated on the left).

(D) HMT assays with indicated factors, chromatin substrates, and 3H-SAM (autoradiographic analysis).

(E) H3K4 methylation state-specific HMT assays with indicated factors (immunoblot analysis with antibodies indicated on the left).

(F) HAT and HMT assays with indicated factors and chromatin templates assembled with the mutant H3 and H4 histones indicated at the top and detailed in Figure S3 (immunoblot analysis with antibodies indicated on the right).

(G–I) MS analysis of p53- and p300-dependent methylation products generated by SET1C and hMLL3/4C (f:PA1C) on chromatin with WT histones (as in E; G shows raw data values, and H shows plotted values) or by SET1C on chromatin with WT or mutant histone (H3KR or H4KR) octamers (as in F; I shows plotted values).

See also Figures S1, S2, S3, S4, and S6.