Fig. 4.
Soluble uPAR shed from mouse OSCC cells induce migration. a-b. Western blot analysis of uPAR protein levels in concentrated conditioned medium from AT84-EV and AT84-uPAR cells. a. Conditioned medium from cells cultured in SFM or SFM containing 10 nM plasminogen (plg) as indicated. The conditioned medium was concentrated from 2 ml to 110 μl for both samples, and deglycosylated using PNGase F. b. Conditioned medium from cells cultured in SFM or in FBSM was concentrated from 2 ml to 350 μl for each sample. A whole cell lysate sample of AT84-uPAR cells was used as a positive control. The two indicated FBSM samples are replicates. Glycosylated suPAR is indicated as suPARglc. c. Non-concentrated conditioned medium from AT84-EV (EV medium) and AT84-uPAR cells (uPAR-medium) was used as an attractant for the AT84-EV cells in a real-time cell migration analysis using the xCELLigence system. N = 3, where each experiment had two technical replicates, and the error bars represent the standard error of mean (±SEM) based on three separate experiments