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Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: Targeting the cross-talk between Urokinase receptor and Formyl peptide receptor type 1 to prevent invasion and trans-endothelial migration of melanoma cells

Fig. 1

Comparison of migration, matrigel invasion and trans-endothelial migration ability of uPAR expressing A375 and uPAR lacking M14 melanoma cells. a Representative images of human A375 and M14 melanoma cells immune-stained with R4 anti-uPAR monoclonal antibody and visualized by a fluorescence inverted microscope. Nuclei were stained blue with DAPI. Scale bar: 5 μm. Original magnification: 1000 x. b Whole cell lysates (40 μg/sample) from A375 and M14 cells were resolved on a 10% SDS-PAGE followed by Western blotting with 1 μg/mL R4 anti-uPAR monoclonal antibody or 0.2 μg/mL anti-GAPDH polyclonal antibody as loading control. c Quantitative Real-Time PCR of uPAR in A375 and M14 melanoma cell lines. Results are the mean ± SD of three different experiments. **: p < 0.001. d-e Wound healing of A375 and M14 melanoma cells kept in growth medium at 37 °C, under a 5% CO2 atmosphere. One field which includes the scratched path from each dish was selected and scanned sequentially every 30 min for 24 h. Images were recorded at the indicated times by a video-camera connected with a motorized inverted microscope. (Original magnification: 50×). e Square root of the wound area measured at the indicated times. f-g A375 and M14 cell migration (f) or matrigel invasion (g) toward serum-free medium (CTRL), or medium containing 10% FBS as source of chemoattractants, monitored for the indicated times by the xCELLigence system. Data represent mean ± SD from a quadruplicate experiment. h Trans-endothelial migration of A375 and M14 melanoma cells. HUVECs (1 × 104 cells/well) suspended in growth medium, were allowed to grow for 24 h until they formed a confluent monolayer, prior to seeding melanoma cells (1 × 104 cells/well). The breaking of monolayer integrity was monitored in real-time as changes in Cell Index for additional 5 h. Data represent mean ± SD from a quadruplicate experiment

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