Fig 1.
BC2059 efficacy in desmoid cells.
A) Desmoid tumor cells strains and normal dermal fibroblasts (NDF-α) were treated with BC2059 (0-10μM) as indicated. B) Representative desmoid tumor migration and invasion in response to BC2059 treatment assessed with Boyden chamber assays. Error bars represent SD from 3 independent experiments. (*P < 0.05; **P < 0.001).
Table 1.
IC50 values of BC2059-treated normal cells lines and desmoid cell strains.
Fig 2.
Analysis of BC2059-induced cell death in desmoid cell strains.
A) Effects of BC2059 on cell apoptosis were measured by flow cytometry. B) Representative cleaved-caspase 3/7 fluorescent dye images of three desmoid cell strains. Effects of BC2059 on cell caspase-dependent apoptosis were measured using automated IncuCyte imaging. Error bars represent SD from 3 independent experiments. (*P < 0.05; **P < 0.001).
Fig 3.
BC2059 inhibits the binding of β-catenin to TBL-1 resulting in decreased β-catenin activity.
A) TBL1 expression after co-immunoprecipitation (IP) of β-catenin. Lysates from three desmoid cell lines treated with 100 nM of BC2059 or vehicle were subjected to β-catenin immunoprecipitation by magnetic beads using a specific anti-β-catenin antibody. Whole-cell lysates (input) and immunoprecipitants were analyzed by immunoblotting with an anti-TBL1 antibody. B) Cytoplasmic and nuclear expression of β-catenin in 3 DT cell lines after treatment with BC2059 for 96h. C) mRNA levels of AXIN2 measured by quantitative real-time PCR after 48h treatment with BC2059. Error bars represent SD from 3 independent experiments. (*P < 0.05; **P < 0.001).
Fig 4.
BC2059 efficacy in desmoid explant cell culture.
A) Cell survival analysis (alamar blue) of S45F-mutated DT tissue treated ex vivo with 100 nM of BC2059 for 96h. B) Representative cytoplasmic and nuclear expression of β-catenin in S45F-mutated DT tissue treated ex vivo with 100 nM of BC2059 for 96h. C) mRNA levels of AXIN2 measured by quantitative real-time PCR in S45F-mutated DT tissue treated ex vivo with BC2059 for 48h. D) Representative protein levels of Axin2 in S45F-mutated DT tissue treated ex vivo with increasing doses of BC2059 after 96h. Error bars represent SD from 3 independent experiments. (*P < 0.05; **P < 0.001).