Fig. 3

Inhibition of uPA-activity reduces migration and invasion in an uPAR-dependent manner. a-e. Real-time cell analysis of proliferation and migration of the AT84-EV and AT84-uPAR cells using the xCELLigence system. N = 3, where each experiment had two technical replicates. The y-axis shows the arbitrary “cell index” value based on cell adhesion induced impedance. Error bars show the ±SD. Student T-test; * p < 0.05, ** p < 0.005, *** p < 0.001. Fold difference was calculated using the mean values. a. One representative proliferation experiment. The standard deviation is based on the two technical replicates. The two vertical black lines show the 24- and 48 h points. Purple line = AT84-uPAR, green line = AT84-EV. Flat purple line = negative control; no cells added. b. Cell index values collected from three separate proliferation experiments at 24- and 48 h. c. Cell index values collected from three separate migration experiments at 24- and 48 h. d. Proliferation of AT84-uPAR cells treated with 10 μM BC11, 10 nM rmPAI-1 or 2 ng/ml TGF-β1 for 48 h. Controls received no additives. e. Migration of AT84-uPAR cells treated with 10 μM BC11, 10 nM rmPAI-1 or 2 ng/ml TGF-β1 for 48 h. Controls received no additives. f. AT84-EV and AT84-uPAR cells invaded the tissue of the leiomyoma invasion model for 7 days with or without the uPA-inhibitor BC11 hydrobromide present (+BC11). Controls received no additives (Ctrl), N = 3. Invasion depth was determined using at least 6 measurements per tissue section. The bars show the mean values of the three discs and the error bars show the ±SEM (N = 3). g. Representative tissue sections of invading AT84-uPAR cells were immunohistochemically stained for uPAR (right panels). Positive uPAR staining is seen as brown colour, counterstained with haematoxylin. Images were recorded at 20 x magnification