Author response image 1. TRIP-Br1-FLAG was detected by TRIP-Br1 antibody.
HeLa cells were seeded in a 60-mm dish the day before the transfection. 5 ug TRIP-Br1-FLAG or control vector were transfected into the HeLa cells. After 48 h culture, cells were lysed by RIPA buffer and the cell lysate was boiled at 95°C for 10 min and added with 200 mM DTT before subject to western blotting by anti-FLAG and anti-TRIP-Br1 antibodies separately. 12% gel was used for better separation of endogenous (arrowhead) and FLAG-tagged TRIP-Br1.