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. 2014 Aug 26;9(8):e105929. doi: 10.1371/journal.pone.0105929

Figure 6. Leiomyoma conditioned medium induced uPAR expression.

Figure 6

Analysis of uPAR expression induced by the LCM or purified ECM proteins in cultured uPAR knock-down cells. All Western blots were performed on whole cell lysates, and uPAR was detected using the polyclonal anti-murine uPAR antibody (AF534). A: Cells were either cultured in LCM (LM) or serum free medium (SF) for 24 hours. Cells were harvested with sample buffer and re-probing for β-actin was used as a loading control. B: uPAR1-NT cells were seeded on different ECM protein substrates, incubated for 24 hours and harvested using RIPA buffer. 7.5 µg of total protein was loaded per lane. Equal loading was verified by re-probing for β-actin. The poloxamer pluronic was used as a no-adhesion control. Col I = Collagen I, Vn = Vitronectin, Fn = Fibronectin, Lm = Laminin, ECL = Entactin, Collagen, Laminin, and FBS = Foetal Bovine Serum. C: Cells cultured in LCM (LM) or serum free medium (SF) for 24 hours were harvested using sample buffer and deglycosylated by PNGase F treatment (+) as indicated. Re-probing for β-actin was used as a loading control. The three bands detected by the anti-uPAR antibody are labelled 1, 2, and 3, respectively.