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Fig. 5. Both uPA-uPAR and uPAR association with β1 integrins are critical to lung cancer cell invasion. (A) Matrigel invasion of H1299 cells. Control, uPAR knockdown (shu), WT, H, D, or HD uPAR expressing H1299 cells were seeded into 24-well Transwell units (1x10⁵ cells/well in triplicate) and allowed to invade Matrigel for 24 hours. The cells that had passed through the filter and attached to the bottom of the filter coated with fibronectin were fixed and stained with Giemsa. The stained cells on the membrane were extracted with 10% acetic acid. Cell invasiveness was evaluated by measuring OD_{595 nm}. (B) Effects of inhibitors on H1299 cell invasion. H1299 cells were pretreated with antibodies (10 µg/ml): 394, uPA activity neutralizing antibody; ATN617, uPAR mAb blocking uPA binding, or control IgG; peptides (100 µM): β1P1 or scβ1P1; inhibitors: GM6001 (0.1 µM), PD98059 (10 µM) or AG1478 (1 µM). The invasion assay was carried out as described above. The results are expressed as a percentage of inhibition observed in the no treatment control. All data are representative of three independent experiments carried out in triplicate.

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