Figure 3: Dkk-3 inhibits cellular motility, anchorage-independent growth in soft agar and cellular viability. (a) Wound assay—photographs of scratch wounds at 0 and 12 hours after wounds were made. After 12 hours of incubation, under the fluorescence microscope, the Dkk-3-143B-transfected OS cells showed less migratory effect compared to the control vector. (b) and (c), Anchorage-independent colony formation assay showed decreased amount of colonies formed by Dkk-3-143B-transfected OS cells (both mixture of all Dkk-3 cells and a single Dkk-3 no. 12 plate) compared to transfected vector control 143B cells. (b) Photograph of soft agar colonies at 18 d after cell seeding. (c) Columns—mean number of colonies; bars—SEM. (d) MTT assay at 72 hours showed less cell viability for Dkk-3-transfected 143B cells (mix, individual cell plates nos. 12 and 18) compared to control vector. Experiments were replicated thrice. (e) Dkk-3-transfected 143B OS cells reduced TCF4 transcriptional activity compared with vector control transfection of 143B cells.