Fig. 4

TRIP12 initiates K63-linked polyubiquitination of EZH2. A Change of EZH2 ubiquitination level with TRIP12 overexpression in HEK293T cells. MG132 was given at 5 µM for 6 h of treatment 48 h after transfection. B Schematic protocols showing steps of LC–MS/MS analysis. N = 2 individual experiments. C Bar chart showing change of K11-, K48- and K63-linked polyubiquitination with TRIP12 overexpression in HEK293T cells which was identified by LC–MS/MS analysis. D, E Change of EZH2 K11- (D) or K48- (E) linked polyubiquitination with TRIP12 overexpression in HEK293T cells. MG132 was given at 5 µM for 6 h of treatment 48 h after transfection. F Change of EZH2 K63-linked polyubiquitination with TRIP12 overexpression in HEK293T cells. G Change of EZH2 K63-linked polyubiquitination upon siRNA-mediated TRIP12 knockdown in NKYS cells. N = 3 individual experiments for all Co-IPs and representative images are shown