We cotransfected 293T cells with Flag-tagged Tet2 and V5-tagged Neurod2 expression vectors. Forty-eight hours after transfection, cells were lysed, and immunoprecipitation was carried out with anti-Flag, anti-V5, or control immunoglobulin G (IgG) antibodies. The immunoprecipitated proteins were analyzed by Western blotting with anti-Flag or anti-V5 antibodies.

Mouse embryo brain cortices at E15.5 days were used for endogenous co-IP. We used the Active Motif Universal Magnetic Co-IP Kit (no. 54002) to perform the co-IP. Antibodies used were rabbit IgG isotype control (Abcam, ab171870), anti-NEUROD2 (Abcam, ab109406), and anti-TET2 (Proteintech, no. 21207-1-AP). The lysate for the co-IP was precleared using rabbit IgG control antibody for 30 min and gently agitated at 4°C. Immune complexes were collected on Dynabeads Protein G from Invitrogen (10003D) for 30 min. Proteins were separated on SDS gels and subjected to Western blotting.

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