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Histone acetylations occur in a progressive hierarchical order at the single-molecule level. (A) H4K12ac is dependent on H4K16ac; (B) H4K8ac is mostly dependent on H4K12ac; (C) H4K5ac is dependent on H4K8ac. These changes are thus highly proteoform specific, requiring H4K20me2 and a series of prior acetylations. Error bars represent standard error from three biological replicates and all statistics are relative to basal conditions (time 0).
