WNT10A increases colony formation ability.

<p>Colony formation assay was used to evaluate the effect of WNT10A on cell transformation. After 48 h of transfection, 2×10⁴ cells were mixed with 0.3% agarose in 1× complete RPMI and transferred into a coated 6-cm dish, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047649#s2" target="_blank">Materials and Methods</a>. To prolong the effects of transfection, pcDNA-WNT10A-transfected cells were maintained in 200 µg/mL G418, and WNT10A siRNA or β-catenin siRNA knockdown cells were maintained in 10 nM of WNT10A siRNA for 15 days. Forced WNT10A expression in 786-O significantly increased the number of colonies of these cells; cotransfection with β-catenin siRNA reduced the WNT10A promotive effect on colony formation ability. Moreover, each WNT10A siRNA knockdown in Caki-1 reduced the number of colonies of these cells. Bar charts show the number of colonies; significances were analyzed by using the Student’s <i>t</i>-test.