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HUVEC transwell migration assay

HM Haiying Ma
SJ Shenglu Jiang
LD Lili Du
JL Jinfang Liu
XX Xiaoyan Xu
XL Xiaomei Lu
LM Ling Ma
HZ Hua Zhu
JW Jun Wei
YY Yanqiu Yu
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HUVEC vertical migration was assessed by using a transwell migration assay, which was performed with a 24-well transwell insert (8 μm pore size, Corning Costar, USA). Next, 250 μl of different conditioned media containing 20% FBS was added to the lower chamber, and 100 μl of HUVEC suspension containing 3% FBS was seeded into the upper chamber at a density of 2 × 104 cells/well. After being incubated for 18 h, the nonmigratory cells on the upper surface of the membrane were gently removed with cotton swabs. The migrated cells on the lower surface of the membrane were fixed with 4% paraformaldehyde and stained with 0.1% gentian violet. Three to five areas were randomly selected to take pictures and count [33, 34].

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