Analyzing Colon Cancer Cell Migration

Migration assays were performed as previously described^{13 (link), 32 (link)}. Briefly, transwell 12 well plates (8 µm pores, Corning) with fibronectin (Sigma, St. Louis, MO) were seeded with 5 × 10⁵ colon cancer cells per well. Cells were treated with either activin A (10 ng/ml) or TGF-β (10 ng/ml) or follistatin (100 ng/ml) for 30 minutes and TGF-β (10 ng/ml) was added. Cells were then allowed to migrate for 6 hours, stained, and imaged. Images from 5 microscopic fields at the center of each well were counted using ImageJ software^{48 (link)}.

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Staudacher J.J., Bauer J., Jana A., Tian J., Carroll T., Mancinelli G., Özden Ö., Krett N., Guzman G., Kerr D., Grippo P, & Jung B. (2017). Activin signaling is an essential component of the TGF-β induced pro-metastatic phenotype in colorectal cancer. Scientific Reports, 7, 5569.

Publication 2017

transwell 12 well plates fibronectin

Activin a Cells Colon cancer Fibronectin Follistatin Microscopic Migration assays Tgf β

Corresponding Organization :

Other organizations : University of Illinois at Chicago, University of Oxford

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Variable analysis

independent variables

Activin A (10 ng/ml)
TGF-β (10 ng/ml)
Follistatin (100 ng/ml)

dependent variables

Number of migrated colon cancer cells

control variables

Transwell 12 well plates (8 µm pores, Corning)
Fibronectin (Sigma, St. Louis, MO)
Seeding density of 5 × 10^5 colon cancer cells per well
Migration duration of 6 hours

controls

Positive control: TGF-β (10 ng/ml) alone
Negative control: Not explicitly mentioned

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