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. Author manuscript; available in PMC: 2010 Nov 20.
Published in final edited form as: ACS Chem Biol. 2009 Nov 20;4(11):958–968. doi: 10.1021/cb9002255

Figure 6. Investigation of ubiquitylation/methylation stoichiometry.

Figure 6

a, Octamers formed with varying ratios or uH2BG76A and H2B were separated on a 15% SDS-polyacrylamide gel and stained with Coomassie. b, Nucleosomes formed with octamers in a, were methylated with Dot1L and 3H SAM. Assay samples were separated by native electrophoresis and stained with ethidium bromide (bottom panel) followed by probing for 3H methyl incorporation by fluorography (top panel). c, Quantification of assay samples by filter-binding followed by liquid scintillation counting, plotted relative to the fully ubiquitylated sample. The blue line (rel. CPM = (fub); where fub is the fraction of uH2BG76A) represents the case where each uH2BG76A stimulates methylation of only one H3K79 side chain in the same nucleosome. The red line (rel. CPM = −(fub)2 + 2(fub)) represents a case where each uH2BG76A stimulates methylation of both H3K79 side chains in the same nucleosome. Error bars represent one s.d. (n = 4).