Fig. 1: Residue S583 of human SUZ12 is phosphorylated.

a Domain structure of SUZ12. Structurally characterized SUZ12 domains are represented by gray blocks except that the VEFS domain included in the current study is colored in green. The PDS loop harboring S583 is highlighted in orange with the amino acid sequence shown above. b Peptides identified for SUZ12 by LC-MS/MS which contain S583. Peptide sequence, modifications, number of peptide spectrum matches (PSMs), and peptide abundance are listed. The phosphorylated residue that was unambiguously assigned is shown in orange. The percentage of phosphorylation was calculated based on a comparison of the abundances of the phosphorylated and unphosphorylated peptides. c Dot blot. Apo and phosphorylated peptides were applied on a nitrocellulose membrane with a serial dilution. Phospho-specific reactivity of the developed anti-SUZ12S583p antibody was analyzed. A representative of three independent experiments is shown. d Effect of λ phosphatase treatment and serine mutation on ectopically expressed PRC2-5m, EZH2–EED–SUZ12–RBBP4–AEBP2. The total amount of PRC2-5m is indicated by anti-SUZ12 signals. S583 phosphorylation level is indicated by signals of the anti-SUZ12S583p antibody developed in this study (uncropped gel images of this figure are shown in Supplementary Fig. 16). A representative of three independent experiments is shown. e Levels of S583 phosphorylation in stem cells and cancer cells. Anti-SUZ12S583p signals were generated using immunoprecipitates of anti-SUZ12 antibody to avoid a non-relevant contaminating band (also see Supplementary Fig. 2). A representative of three independent experiments is shown. Source data are provided as a Source Data file.