Fig. 7

PDPN silencing alters the expression and activity of matrix metalloproteinases (MMPs) in TPC1 and BcPAP cells. a. The pattern of expression of MMPs mRNA levels in TPC1 and BcPAP cells. b. RT-qPCR analysis of MMP mRNA levels in 48 h after siPDPN transfection compared to those in non-transfected cells. The results were normalized to 18S RNA levels. The bars represent the average fold expression. Data are presented as means ± standard errors of the mean (SEM) of results from at least three independent experiments *P < 0.05. c. Western blot analysis of MMP expression levels 48 h after PDPN silencing. Total protein extracts were separated by SDS-PAGE, transferred onto a nitrocellulose membrane, and probed with specific antibodies. The results are representative of three independent experiments. d. Immunofluorescent staining for MMP-2 and MMP-9 in thyroid cancer cell lines transfected with siPDPN or control siNeg. Cells were fixed with 4% PFA/PBS, permeabilized with 0.1% Triton X-100, and stained with an anti-MMP-2 or anti-MMP-9 monoclonal antibody followed by the DyLight549-conjugated secondary antibody (red), and counterstained with DAPI (blue). Confocal microscopy magnification: 630x. e. Gelatin and collagen zymography for TPC1 and BcPAP cell lines after PDPN silencing