Matriptase expression enhances spheroid adhesion, mesothelial clearance, and migration/invasion. (A) Representative image of spheroid adhesion to mesothelial monolayers. NCI/ADR-Res Vec and Mat spheroids were formed on agarose hydrogel and seeded onto confluent LP9 monolayers (formed on thin collagen), allowed to adhere for 1 h, washed, and adherent spheroids visualized by EVOS; representative stitched images taken at 4× magnification of whole-well scans are shown (scale bars represent 1.4 mm). A quantitative analysis of three independent experiments performed in triplicate is provided in Fig. 3 A. (B) Mesothelial clearance. NCI/ADR-Res Vec or Mat spheroids were formed on agarose hydrogel, seeded onto confluent, fluorescently labeled LP9 monolayers (calcein-AM), and mesothelial clearance was visualized up to 5 h by EVOS; representative images are shown at 4× magnification, scale bars represent 650 μm. (C) Migration and invasion. SKOV3 Vec and Mat cells were seeded onto uncoated or Matrigel-coated transwells, allowed to migrate or invade for 16–24 h, stained by Kwik-Diff, and quantified by ImageJ. Data represents average migration/invasion relative to Vec from three independent experiments performed in triplicate (error bars are ± SEM; **P < 0.01, two-tailed unpaired Student’s t test).