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Fig. 5. Podoplanin-induced EMT is associated with increased migratory and invasive abilities. (A) In vitro wound-healing assay. Confluent cell cultures were scratched with a pipette tip to produce a wound and analyzed by time-lapse video microscopy. Images of the wounded area immediately (0 hours) and 12 hours after the incision was made are shown. Arrows in P
CT indicate individual cells moving into the wound as fibroblasts. An animated sequence of these data is shown in supplementary material Movie 1. In the bottom panel a graph shows quantification of cell migration from three independent experiments. **P<0.01; ***P<0.001 vs control (EGFP) cells. (B) Matrigel invasion assay. Cell transfectants expressing EGFP-tagged wild-type and mutant podoplanin proteins were seeded on a 24-well Matrigel invasion chamber. FBS (5%) was used as a chemoattractant. Fluorescent cells that invaded the Matrigel-coated filter after 24 hours were detected by confocal microscopy. Images were acquired each 2 µm along the z-axis and quantified by fluorescence intensity profiles. Vertical (x-z) sections of the filter are shown below. Results are representative of two experiments.
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