Activation of Wnt/β-catenin signaling pathway in spheroids/colonospheres. A. Representative photomicrographs of HT-29 parental and colonospheres-derived cells depicting the membrane localization of β-catenin by immunofluorescence staining. B. The comparative expression of different members of Wnt/β-catenin signaling pathway in parental cells (referred to as 'P') and colonospheres-derived cells (referred to as 'S'). The Wnt/β-catenin signaling is constitutively activated in colonosphere-derived cells, but not in parental cells as shown by the expression of various proteins: β-catenin, phosphorylated β-catenin, axin-1, phosphorylated GSK-3-β along with important downstream targets: c-myc and cyclin-D1. The numbers represent percent of corresponding control normalized to β-actin.