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. 2016 May 23;113(23):E3213–E3220. doi: 10.1073/pnas.1600509113

Fig. S2.

Fig. S2.

HIRA and UNB1 are O-GlcNAcylated. (A) HIRA is O-GlcNAcylated. Flag-tagged HIRA and Myc-tagged OGT were expressed in HEK293T cells. Flag-tagged HIRA and associated proteins were immunoprecipitated with FLAG M2 beads. Immunoprecipitated (IP) proteins and total cell extracts (input) were analyzed by Western blot analysis using the indicated antibodies. (B and C) UBN1 is O-GlcNAcylated by OGT. GFP-tagged UBN1 and Myc-tagged OGT were expressed in HEK293T. UBN1-GFP was immunoprecipitated, and associated proteins were analyzed by Western blotting using the indicated antibodies (B) or by mass spectrometry (C). (D) Quantification of modified and unmodified peptide of HIRA. Selected ion chromatograms of 1,061.01 [M+2H]2+ and 1,162.55 [M+2H]2+ that correspond to the unmodified and the O-GlcNac modified, respectively, are shown (Left). The peak area of the O-GlcNAc–modified peptide ions is ∼4% of the unmodified ion peak area (Right).