Fig. 6.

S100A4 knockdown by siRNA leads to MMP2 down-regulation. Unsorted OCTT2 cells were transfected either with nontargeting siRNA (siN) or siS100A4 oligos si1 and si2, respectively. Knockdown of S100A4 was quantitated by qPCR (A, left graph), Western blot of whole cell lysates (WCL) (B), MRM analysis (C) and immunofluorescence (D) 72 h after transfection. Down-regulation of S100A4 led to concomitant decrease in MMP2 expression, as measured by qPCR (A, right graph) Western blot (B), and MRM (C). For the MRM analysis a third siS100A4 oligo was included (si3). MRM confirmed that the amount of soluble S100A4 was significantly decreased in the siS100A4 samples compared with the siN oligo, and MMP2 was significantly decreased with si2 and si3. Representative zymographic analysis shows decreased proteolytic activity in CM collected from si3-transfected cells. n = 3, *p < 0.05.