Co-immunoprecipitation (co-IP) assay
Co-IP assay was performed using the Pierce co-IP kit (Thermo scientific 26149), as per the manufacturer's instructions. The total lysate extracted from HCC cells expressing OGT-FLAG was IP with FLAG Ab or EZH2 Ab, after which Western blotting was performed using the designated antibodies (Abs). The total lysate of HCC cells treated with TMG was IP with IgG or O-GlcNAc Ab (RL2), followed by Western blotting with EZH2 Ab, and Ig heavy chain (H chain) was used as loading control. The antibodies used included the following: EZH2 (5246, CST); O-GlcNAc (ab2739, abcam); Flag (SAB4200071, Sigma-Aldrich); IgG (NI01-100UG, Millipore).

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