Icrt14

Cells and clinical information from MCL patients described in this manuscript (Additional file 4: Table S2) were collected and published with the written informed consent of each patient under The University of Texas MD Anderson Cancer Center IRB-approved clinical protocol LAB08-0190 for use of human tissues.
The following agents were tested: Wnt inhibitors XAV939 (Selleck Chemicals, Houston, TX), iCRT14 (R&D Systems, Minneapolis, MN), CCT036477, PKF118-310 (Sigma-Aldrich, St. Louis, MO), IWP2, IWR1-endo, and IWR1-exo (Santa Cruz Biotechnologies, Santa Cruz, CA); Hedgehog inhibitors GANT61 (R&D Systems, Minneapolis, MN), LDE225 and Cyclopamine (both from Selleck Chemicals, Houston, TX); Notch inhibitor RO4929097 (Selleck Chemicals, Houston, TX).

Luciferase activity was assessed 48–72h post-transfection. Experiments were performed in 24-well plates with reduced serum (2.5%). For Wnt pathway blocking experiments, cells were cultured in reduced serum (2.5%) media containing either 0.1μg/mL Dkk1 or 50μM iCRT14 (R&D Systems). At each time point media samples were taken and analysed for secreted Gaussia luciferase activity using a luciferase flash assay kit (Thermo Pierce) on a Biotek Synergy 2 plate reader with automatic injection system controlled with Gen5 software. Luciferase activity was normalised to the total protein content of the cell lysates obtained using luciferase assay cell lysis buffer (Thermo Pierce) taken at the experimental end-point. Fold changes for each treatment were calculated relative to the cells only control.