Figure 4. SFRP3 can inhibit Wnt5a signaling.

A, Topflash reporter assay. A2058 cells transiently transfected with Topflash and Fopflash plasmids. Where appropriate, cells were pre-treated with 1 µg/ml rSFRP3 or carrier, and after 24 h cells were treated with the indicated combinations of Wnt ligands (0.2 µg/ml rWnt5a and 0.05 µg/ml rWnt3a) as indicated for an additional 24 h. The image is representative of 5 independent experiments and the error bars represent SD within one experiment. B, Measurement of intracellular Ca²⁺ signaling. rWnt5a (0.1 µg/ml, addition indicated by arrows) was added to A2058 cells pre-treated with carrier (upper panel) or 1 µg/ml rSFRP3 (lower panel). Traces are representative images of 4 separate experiments. C, ΔCa²⁺ ratio values from A2058 cells pre-treated with either carrier or 1 µg/ml rSFRP3 and then stimulated with 0.1 µg/ml rWnt5a. Error bars represent SD (n = 4). ** = p<0,01.