ML141 (CID-2950007) is an effective, specific and reversible non-competitive inhibitor of Rho family GTPase cdc42 (IC50: 200 nM).

CDC42:200 nM

在负荷MDA-MB 231源肿瘤的NOD/SCID小鼠体内, ML141（1 mg/day,i.p.）通过抑制Cdc42可使TMX抑制MDA-MB 231衍生的肿瘤的生长.此外, ML141（10 mg/kg,i.p.）可增强小鼠体内粒细胞集落刺激因子诱导的造血干细胞和祖细胞动员.

ML141对二甲双胍诱导的神经母细胞瘤凋亡损害具有明显的保护作用。通过对细胞死亡/分裂的诱导/抑制,ML141可使咖啡因抑制细胞生长的能力增强。ML141还可剂量依赖性减少肺炎克雷伯菌入侵。

Equilibrium binding assay : Wild-type GST-Cdc42 (4 μM) is bound to GSH-beads overnight at 4°C. Cdc42 on GSH-beads is depleted of nucleotide by incubating with 10 mM EDTA containing buffer for 20 min at 30°C, washing twice with NP- HPS buffer, then re-suspended in the same buffer containing 1 mM EDTA/or 1 mM MgCl2, 1 mM DTT and 0.1% BSA. Cdc42 unbound sites are blocked by incubation of protein–bead complex for 15 min at RT. Thirty μL of this suspension is incubated with 20 mM inhibitor for 3 min at RT and added 30 μL of various concentrations of ice cold BODIPY-FL-GTP. Samples incubated at 4° C for 45 min and binding of fluorescent nucleotide to enzyme is measured using an Accuri flow cytometer. Raw data are exported and plotted using GraphPad Prism software.

Cells are incubated with 500 nM Calcein-AM and 1 µM PI for 15 min, after which live cells and dead cells (represented by positivity of Calcein-AM and PI staining, respectively) are counted utilizing the adherent cell Celigo™ cytometer. (Only for Reference)