Human lymphocytes or CD4+ T cells were cultured in RPMI 1640 or Iscove’s modified Dulbecco’s medium with KnockOut Serum Replacement (all from Life Technologies). GT13072 or GT14829 was added for 10 min. Proteins were extracted with radioimmunoprecipitation assay buffer supplemented with Protease Inhibitor Cocktail and Phosphatase Inhibitor Cocktail according to the manufacturer’s instructions (all from Sigma-Aldrich). Protein (20 μg) was loaded on a NuPAGE Novex 4-12% Bis-Tris gel and separated using the NuPAGE Bis-Tris Electrophoresis System (Invitrogen). Proteins were transferred onto a 0.2-μm polyvinylidene difluoride membrane (Amersham). The antibodies used are listed in table S4. Bound antibodies were identified with the ECL Plus Western Blotting Detection Kit (GE Healthcare).

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