Co-immunoprecipitation (CO-IP) assays using transfected HEK293T cells

HEK293T cells were transfected with the indicated plasmids using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions for 24 h and lysed in cell lysis buffer (Cell Signaling Technology) supplemented with Protease Inhibitor Cocktail (MCE). Cell lysates were immunoprecipitated overnight at 4°C using anti-HA magnetic beads (Thermo Fisher Scientific, 88836) or anti-MYC magnetic beads (Thermo Fisher Scientific, 88843). The magnetic beads were washed three times with TENT buffer. The precipitates were immunoblotted with HA and MYC antibodies.