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Transwell assay

SD Shuhong Dai
NL Ning Li
MZ Ming Zhou
YY Yue Yuan
DY Ding Yue
TL Tao Li
XZ Xiaowei Zhang
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To determine cell invasion ability, the cells (8 × 102 cells) were resuspended in serum-free medium and seeded onto the upper chamber of a Matrigel-coated Transwell insert (EMD Millipore). Complete medium supplemented with 10% FBS was added to the lower chamber. After 24 h, the upper surface of the membrane was wiped with a cotton swab, and the cells attached to the lower surface were fixed with 4% formaldehyde for 10 min at room temperature and stained with 1% DAPI (4′, 6-diamidino-2-phenylin-dole) solution for 10 min. The invaded cells were observed and counted under a light microscope. The procedure was carried out as described previously32.

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