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. 2012 Jun 15;125(12):2954–2964. doi: 10.1242/jcs.101592

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© 2012. Published by The Company of Biologists Ltd

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Fig. 2. — H4 K12ac and H3 K4me3 are present in Htz1p-containing mononucleosomes, as shown by PIE-FRET and FCS. (A) PIE-FRET efficiency histograms of interactions of Htz1–HA plus H4 K12ac of mononucleosomes isolated from wild-type yeast (left panel) or H4 K12R mutants (middle panel) expressing HA–Htz1p. Fluorescence cross-correlation analysis of Htz1–HA plus H4 K12ac of HA–Htz1p mononucleosomes (right panel). (B) PIE-FRET efficiency histograms of interactions of Htz1–HA plus H3 K4me3 of mononucleosomes isolated from wild-type yeast (left panel) or set1Δ mutants (middle panel) expressing HA–Htz1p. Fluorescence cross-correlation of Htz1–HA plus H3 K4me3 of HA–Htz1p mononucleosomes (right panel). Donor: anti-H4 K12ac–FAM-X (A) or anti-H3 K4me3–Alexa-Fluor-488 (B). Acceptor: anti-HA–Alexa-Fluor-647. (C,D) H3 K36me3 is present in H2A mononucleosomes but undetectable in Htz1p mononucleosomes. PIE-FRET efficiency histograms of interactions of H2A (C) or HA–Htz1p (D) with H3 K36me3 in mononucleosomes isolated from wild-type yeast (left panels) or set2Δ mutants (middle panels) expressing HA–Htz1p. Fluorescence cross-correlation analysis of H3 K36me3 plus H2A of wild-type yeast mononucleosome (C, right panel). Donor: anti-H3 K36me3–Alexa-Fluor-488. Acceptor: anti-H2A–Alexa-Fluor-647 (C) or anti-HA–Alexa-Fluor-647 (D). (E) H4 K12ac and H3 K4me3 are present in the same nucleosome. PIE-FRET efficiency histogram of interactions between H4 K12ac and H3 K4me3 in mononucleosomes isolated from wild-type yeast (left panel) or set1Δ mutants (middle panel). Fluorescence cross-correlation of H4 K12ac plus H3 K4me3 of wild-type yeast mononucleosomes (right panel). Donor: anti-H3 K4me3–Alexa-Fluor-488. Acceptor: anti-H4 K12ac–Alexa-Fluor-647.