ASZ001 Sant-1 resistant cells were grown to 60% confluency under regular growth media. Cells were treated with DMSO or silibinin (100 μM) and/or Sant-1 (60 μM) and/or GDC-0449 (40 μM) for 48 hrs. At the end, cell lysates were prepared as previously described (3 (link), 34 ), and protein concentration was determined by Bio-Rad DC protein assay kit (Manufacturer’s protocol). Equal protein per sample was resolved on Tris-glycine gels, and transferred onto nitrocellulose membranes. Cell lysates were probed with specific primary antibodies followed by appropriate peroxidase-conjugated secondary antibody and visualized by enhanced chemiluminescence detection system (GE healthcare). To ensure equal protein loading, membranes were stripped and re-probed with appropriate loading control. The bands were scanned with Adobe Photoshop 6.0 (Adobe Systems, San Jose, CA). The band intensity was analysed by using Image J (NCI, USA) tool and presented as fold change to that of their respective controls. The expression in each group was also normalized to their respective loading control.