Fig 1: ITCH may function as an E3 ubiquitin ligase targeting CDK4 for degradation. (A) Relative CDK4 mRNA expression in HCT116 (Vector/HA-CCDC68) and RKO (Vector/HA-CCDC68) cells. Student′s t-test, ns, not significant. (B) CDK4 protein degradation was monitored in HCT116 (CCDC68) and HCT116 (Vector) cells pretreated with MG132 for 5 h followed by CHX for various times. (C) CDK4 protein degradation rates in HCT116 (CCDC68) and HCT116 (Vector) cells. Three independent degradation experiments were performed, and the results were analyzed using ImageJ software and analyzed by Student′s t-test. *P < 0.05. (D) Molecules potentially involved in the regulation of CDK4 protein degradation were predicted by the UbiBrowser database. (E) Venn diagram screening the E3 enzymes involved in the regulation of CDK4 protein degradation. (F) Expression of CDK4, ITCH, BTRC, CDC37, FBXO4, and SOCS7 in HCT116 (Vector/HA-CCDC68) and RKO (Vector/HA-CCDC68) cells.
from Cell Signaling Technology for beta-TrCP (D12C8) Rabbit mAb