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. 2014 Dec;184(12):3384–3393. doi: 10.1016/j.ajpath.2014.08.003

Figure 4.

Figure 4

Urokinase receptor (uPAR) induces transforming growth factor β (TGF-β) expression in MDA-MB breast cancer 468 cells. A: Immunoblot analysis to detect TGF-β and tubulin was performed on cell extracts from empty vector-transfected control (EV) and uPAR-overexpressing MCF7 and MDA-MB 468 cells. B: Conditioned medium from control MDA-MB 468 cells (468/EV) and uPAR-overexpressing MDA-MB 468 cells (468/uPAR) was subjected to enzyme-linked immunosorbent assay (ELISA) to detect TGF-β. C: Control MDA-MB 468 cells (EV) and uPAR-overexpressing MDA-MB 468 cells (uPAR) were transfected with uPA-specific siRNA (40 nmol/L) or NTC siRNA pool (40 nmol/L). After 24 hours, the cells were cultured in serum-free medium for an additional 24 hours. mRNA levels were determined by qPCR for urokinase-type plasminogen activator (uPA) and TGF-β, standardized against levels present in control EV cells transfected with control siRNA. D: Immunoblot analysis to detect TGF-β and tubulin was performed on cell extracts from control and uPAR-overexpressing MDA-MB 468 cells that were transfected with uPA-specific or NTC siRNA. Data are expressed as means ± SEM. n = 3. ∗∗P < 0.01, Student’s t-test.