IWP-2 is a Wnt pathway inhibitor (IC50=27 nM) and an ATP-competitive CK1δ inhibitor (IC50=40 nM for M82FCK1δ). IWP-2 inhibits self-renewal of embryonic stem cells and has been used in stem cell and organoid research.

Wnt:27 nM

方法: 人胃癌细胞 MKN28 用 IWP-2 (5-50 µM) 处理 1-5 天，通过 CellTiter Assay 检测细胞增殖。
结果: 在 MKN28 细胞系中治疗四天后，10-50 µM IWP-2 显著抑制了 MKN28 细胞的增殖。[1]
方法: L-Wnt3A 细胞用 IWP-2 (5 µM) 处理 24 h，通过 Western Blot 检测靶点蛋白表达水平。
结果: IWP-2 阻断了所有检测到的 Wnt 依赖性生化变化，Lrp6 受体和 Dvl2 的磷酸化，以及 β-catenin 的积累。[2]

方法: 为研究对脱发的作用，用 AA 抗氧化剂 (100 mg/mL，局部用于皮肤)、PBMT 和 IWP-2 (20 mg/kg，静脉注射，每两天一次) 治疗 C57/BL6 小鼠。
结果: 在休止期剃须刺激后，PBMT 促进了头发再生。重要的是，使用 AA 和 IWP-2 消除了 PBMT 在体内诱导的 HF 生长速率的差异。[3]

IWP-2 is prepared in DMSO and stored, and then diluted with appropriate medium before use[2]. The human RCC cell lines 786O and Caki-2 (5×103) are seeded into 96-well plates. Cell viability is estimated by MST assay after Caki-2 acells are incubated with ncreasing concentrations of LEF together with 20 μM IWP-2 for 48 h.After treatment, 10 μL MTS is added into each well for 2 h incubation. The absorbance is measured using a model ELX800 Micro Plate Reader at 490 nm. For colony formation assay, Caki-2 cells are trypsinized to single cell suspensions and seeded into fresh 6-well plates at 1000 cells/well. Then cells are incubated with LEF at depicted concentrations for 7 days. Colonies are fixed with absolute methanol for 15 min and then stained with 0.1% crystal violet for 20 min. After washing with PBS three times, the colonies with a diameter over 2 mm are visualized by a digital camera[2].