Extended Data Fig. 1: Post-translational modifications on YTHDFs identified by LC–MS/MS.
From: O-GlcNAcylation determines the translational regulation and phase separation of YTHDF proteins
a) Filtered identified modifications in HEK293T were annotated on the sites (score > 100 for all modifications and diagnostic peak for O-GlcNAcylation). b, O-GlcNAcylation ratio of each site of YTHDF1 and YTHDF3 by LC–MS/MS analysis in HEK293T calculated with intensities (in blue) and MSMS counts (in orange) of the respective modified and unmodified peptide. (c) Estimated ratio of unmodified, single modified and double modified of YTHDF1 and YTHDF3 by the intensity ratio in (b). The estimated modification level is highly similar to the O-GlcNAcylation stoichiometry analysis in Fig. 1e. (d) O-GlcNAcylation stoichiometry analysis of Flag-tagged MYPT1, CSNK2A1 and CREB1 in HEK293T and HeLa. Arrowheads indicates the O-GlcNAcylation modified proteins. Flag* denoted samples were performed O-GlcNAcylation stoichiometry analysis and immunoblotting by anti-Flag. (e) Quantification result of O-GlcNAcylation level of MYPT1, CSNK2A1 and CREB1 in (d). Data are presented as mean ± s.d. (n = 3 biologically independent repeats). Significance was calculated using two-sided t-test.
