Migration assay was conducted with ibidi® culture insert. Cell was seeded at both side of the insert (35,000 cells in 70 μl medium) and incubated overnight. The insert was removed next day and photographed at 0 and 48 h. The gap between cells was quantified with Image J and presented as percentage of closure compared to 0 h.
The invasion assay was described previously [28 (link)]. Briefly, 100 μl of 80 μg Matrixgel (BD) was previously loaded onto the upper chamber of 24 well transwell (BD, 8 μm pore size) at 37 °C for 2 h and 5 × 104 cells were seeded on the gel (in 200 μl of medium without FBS) and 500 μl of complete medium was added into the lower chamber of the transwell. Cells invaded through transwell were stained after 24 h incubation. Five images were photographed for each transwell under 100X magnification. Cell numbers were counted and calculated.
The invasion assay was described previously [28 (link)]. Briefly, 100 μl of 80 μg Matrixgel (BD) was previously loaded onto the upper chamber of 24 well transwell (BD, 8 μm pore size) at 37 °C for 2 h and 5 × 104 cells were seeded on the gel (in 200 μl of medium without FBS) and 500 μl of complete medium was added into the lower chamber of the transwell. Cells invaded through transwell were stained after 24 h incubation. Five images were photographed for each transwell under 100X magnification. Cell numbers were counted and calculated.
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