A total of 29 WT chromodomains and two CBX1 mutants were expressed as glutathione S-transferase (GST) fusion proteins using pGEX-KG vector in Escherichia coli strain BL21(DE3) based on a previously described protocol (19). A total of 467 unmodified and modified peptides [selected by a bioinformatics pipeline previously developed and described in (19, 20) and in the Supplementary Methods and Materials] were synthesized by Sigma-Aldrich (desalted, mass spectrometry checked). The peptides were then printed as triplets onto glass slides (ArrayIt). The CBX1 mutant was screened against an Active Motif MODified Histone Peptide Array. Binding of the peptide microarray and the chromodomain proteins was performed and analyzed using a previously described protocol [see (19) and Supplementary Methods and Materials for details].

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