Sc 517122

Co-immunoprecipitation (co-IP) was performed according to the manufacturer's instructions (Pierce Co-Immunoprecipitation kit, Thermo Fisher Scientific, Inc.). RCC cells with indicated treatment were used for one immunoprecipitation reaction. Briefly, cells were lysed in a series of buffers and centrifugation steps to obtain lysate supernatant. Indicated antibodies were covalently coupled onto an amine-reactive resin and used to bait the corresponding proteins. Antibodies against QPCT (sc-517122, Santa Cruz Biotechnology, Inc., 1:50) and PIK3CA (4255, Cell Signaling Technology, Inc.; 1:50) were incubated for 12 h at 4°C and then incubated with IRDye 800-conjugated goat anti-rabbit IgG (2095, Li-Cor Biosciences Inc.; 1:1,000) for 1 h in the dark at room temperature, and bands were detected using an Odyssey infrared scanner (Li-Cor).