Phosphorylation Status of Thymidine Kinase 1 Following Antiproliferative Drug Treatment Mediates 3′-Deoxy-3′-[18F]-Fluorothymidine Cellular Retention
Figure 5
Determination of the regulation of TK1 phosphorylation by Cdk1 and Cdk2.
A) Cdk1 and Cdk2 knock-down optimization. Representative western blots, with corresponding densitometry (average of 3 independent experiments) and qPCR analysis. Control refers to non-silenced cells; scramble indicates transfection with non-targeting siRNA. B) Determination of the Cdk responsible for TK1 specific phosphorylation. 5 nM of scramble, Cdk1 or Cdk2 siRNA were used to induce Cdk knock-down for 48 h. Cells were treated O/N with 0.5 µg/ml nocodazole (NOC) following Cdk knock-down. Top two panels represent MnCl2-phos-tag gel; bottom three panels report 12% SDS-PAGE.