Fig. 5
From: CCT8 drives colorectal cancer progression via the RPL4-MDM2-p53 axis and immune modulation
CCT8 breaks the binding with RPL4 and MDM2. (A) HEK293T cells were divided into three groups: Group 1 was transfected with RPL4-Myc, Group 2 was transfected with RPL4-Myc and MDM2-Flag, and Group 3 was transfected with RPL4-Myc, MDM2-Flag and CCT8-Myc. Then, RPL4-Myc, MDM2-Flag and CCT8-Myc antibody markers were used for each group. The transfection efficiency was verified by western blotting. (B) MDM2-Flag was pulled down by anti-Flag beads. MDM2-Flag and RPL4-Myc were examined with anti-Myc and anti-Flag antibodies. Western blotting showed reduced binding of MDM2 and RPL4 after transfection with CCT8. (C) Group 1 was transfected with p53-Flag and Ub-HA, Group 2 was transfected with p53-Flag, Ub-HA and RPL4-Myc, and Group 3 was transfected with p53-Flag, Ub-HA, RPL4-Myc and CCT8-Myc. Flag-p53 was pulled down by anti-Flag beads and assessed with anti-HA antibodies. Western blotting demonstrated that CCT8 enhances MDM2-mediated p53 proteasomal degradation. Experiments including gels and blots were repeated three times