Figure 5.

siRNA-based knock down of PAX2 and WNT5A genes. Two locations within PAX2 and WNT5A genes were specifically targeted to knock down their expression using commercialized siRNA (Qiagen). A nonspecific siRNA was used as a negative control where protein expression is not altered in treated samples compared to nontreated samples (A). It is noteworthy that WNT5A expression is weak when compared to the PAX2 expression in the WiT49 WT cell line. Both PAX2-siRNA₁ and PAX2-siRNA₂ were efficient to reduce drastically PAX2 expression (B), whereas WNT5A expression was clearly detectable in the same samples (D). Culture medium where WiT49 cells were propagated and treated with WNT5A-siRNA₂ and WNT5A-siRNA⁶ was concentrated and subjected to Western blot analysis using an anti-Wnt5A antibody. Like PAX2-siRNA, both WNT5A-siRNA₂ and WNT5A-siRNA₆ reduced the WNT5A expression (C). The presence of PAX2 expression was detected in these WNT5A knocked down samples (E).