Active site-targeted covalent irreversible inhibitors of USP7 impair the functions of Foxp3+ T-regulatory cells by promoting ubiquitination of Tip60
Fig 6
Selective USP7 inhibitor P217564 regulates the stability of USP7.
(A) P217564 decreases the USP7 level in a dose-dependent manner. Jurkat cells were treated with DMSO or P217564 at indicated concentrations for 24 hours; crude cell extracts were subjected to SDS-PAGE electrophoresis, followed by IB with indicated antibodies. *n.s indicates non-specific band. Relative quantitation of band intensity is given. (B) P217564 decreases the half-life of USP7. HCT116 cells were treated with DMSO or P217564 in the presence of 50ug/mL cycloheximide (CHX), and the levels of USP7 and HDM2 were analyzed using western blotting at indicated time points. The IB of Actin was used as the loading control.