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. Author manuscript; available in PMC: 2020 Mar 4.
Published in final edited form as: Nature. 2019 Sep 4;573(7773):281–286. doi: 10.1038/s41586-019-1534-3

Extended Data Figure 6: PWWP domain of DNMT3A preferentially binds to H3K36me2.

Extended Data Figure 6:

a) The dCypher approach to interrogate chromatin readers (see Methods). Biotinylatednucleosomes are immobilized on Streptavidin donor beads and GST-tagged DNMT3APWWP on glutathione AlphaLISA acceptor beads. Laser excitation of the donor generates singlet oxygen that diffuses to activate emission from the acceptor: fluorescence counts are directly proportional to the amount of donor-acceptor bridged by the nucleosome-reader interaction.

b) Quantification of AlphaLISA counts for isolated GST-DNMT3APWWP interaction titratedagainst H3K36-modified nucleosomes (me0/me1/me2/me3) from mean of two replicates.

c) ITC titration and fitting curves of human DNMT3A PWWP domain with H3.3(1-42)K36-modified peptides (me0/me1/me2/me3).

d) ITC titration and fitting curves of human DNMT3A PWWP domain with H3.1(1-42)K36-modified peptides (me0/me1/me2/me3) .