Cell cycle–specific AChE-seq studies of macroH2A incorporation in S/G2 and G1 phases of the cell cycle. (A) The time line of synchronizing, blocking, and harvesting of cells. (B) The experimental overview of AChE. (C) MacroH2A enrichment “peaks” overlap substantially between macroH2A newly incorporated in S/G2 and in G1. (D) MacroH2A is enriched before and after cell division in Giemsa-positive bands, specifically those categorized as gpos100 in the UC Santa Cruz Genome Browser. The results of permutation tests demonstrate enrichment only for the most heterochromatic cytogenetic bands (P < 0.01). (E) AChE-qPCR of loci predicted from the AChE-seq results to be positive and negative validates these genome-wide studies. Pull-down without SNAP-biotin treatment was used as negative controls (mock). The graph shows means and standard deviations from three independent experiments.