Figure 4

Wnt-dependent role of ROR2 in colon cancer cells. (A) Restoration of ROR2 inhibits β-catenin/TCF transcription activity. DLD1 colon cancer cells were transiently transfected with TOPflash or FOPflash reporters alone (white) or in the presence of different amounts of an ROR2 expression construct (black). Cells were lysed 48 h post-transfection and Firefly and Renilla luciferase activities measured. Results are shown as means ± SD of triplicate samples from one representative experiment. (B) WB showing ROR2 expression in DLD1 mock-and ROR2-transfected cells (left). MTT analysis of viability of the same cells in the presence of anti-WNT5A antibody (2 μg/ml; upper right) or recombinant WNT5A (200 ng/ml; lower right). Treatment was renewed daily. Data are expressed as the percent MTT signal relative to the untreated sample at the same time point. (C) Immunofluorescence staining of β-catenin and α-lamin B in DLD1 mock-and ROR2-transfected cells after 2 h treatment with recombinant WNT5A (200 ng/ml) or anti-WNT5A antibody (2 μg/ml) or untreated (CTR). (D) TOPflash assay (as in A) of the same cells after 72 h treatment (as in B) with recombinant WNT5A or anti-WNT5A antibody.