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, CK1δ:40 nM

METHODS: Human gastric cancer cells MKN28 were treated with IWP-2 (5-50 µM) for 1-5 days, and cell proliferation was detected by CellTiter Assay.
RESULTS: After four days of treatment in MKN28 cell line, 10-50 µM IWP-2 significantly inhibited the proliferation of MKN28 cells. [1]
METHODS: L-Wnt3A cells were treated with IWP-2 (5 µM) for 24 h, and the expression levels of target proteins were detected by Western Blot.
RESULTS: IWP-2 blocked all detected Wnt-dependent biochemical changes, phosphorylation of Lrp6 receptor and Dvl2, and accumulation of β-catenin. [2]

METHODS: To study the effects on hair loss, C57/BL6 mice were treated with AA antioxidant (100 mg/mL, topically applied to the skin), PBMT, and IWP-2 (20 mg/kg, i.v., every two days).
RESULTS: PBMT promoted hair regrowth after resting shaving stimulation. Importantly, the use of AA and IWP-2 eliminated the differences in HF growth rates induced by PBMT in vivo. [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].

DMSO: 1 mg/mL (2.14 mM), Sonication is recommended.