CK-636 (CK-0944636), an Arp2/3 complex inhibitor, can inhibit actin polymerization induced by the human (IC50: 4 μM), fission yeast (IC50: 24 μM) and bovine (IC50: 32 μM) Arp2/3 complex, respectively.

Arp2/3 complex (human):4 μM, Arp2/3 complex (fission yeast):24 μM, Arp2/3 complex (bovine):32 μM

通过抑制迁移T细胞前缘板状伪足的形成,CK-636可减慢迁移速度。在活细胞中,CK-636对Arp2/3复合体介导的肌动蛋白聚合有抑制作用,在感染的SKOV3细胞中,CK-63可使肌动蛋白丝的形成减少。此外,CK-636处理的大部分T细胞在通过字型测试实验时会出现一个转角,然后陷入Z字型的界面上。

HDAC inhibition assays: HDAC inhibition assays are performed by Reaction Biology Corp. using isolated human, recombinant full2length HDAC1 and -6 from a baculovirus expression system in Sf9 cells. An acetylated fluorogenic peptide,RHKKAc, derived from residues 379-382 of p53 is used as substrate. The reaction buffer is made up of 50 mM Tris-HCl pH 8.0, 127 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 1 mg/mL BSA, and a final concentration of 1% DMSO. Compounds are delivered in DMSO and delivered to enzyme mixture with preincubation of 5-10 min followed by substrate addition and incubation for 2 h at 30°C. Trichostatin A and developer are added to quench the reaction and generate fluorescence, respectively. Dose-response curves are generated starting at 30 μM compound with three-fold serial dilutions to generate a 10-dose plot. IC50 values are then generated from the resulting plots, and the values expressed are the average of duplicate trials ± standard deviation.