The cell invasion assay was performed using six-well Matrigel invasion chambers with 8-μm pore inserts (BD Biosciences). Cells (5 × 104) cells were seeded into the upper chambers. The lower chambers were filled with serum-free medium. After 24 h, the cells in the upper chamber were treated with HuR-FNP in serum-free medium. After 6 h of transfection, the culture medium in the upper chamber was replaced with 2% serum-containing medium with or without AMD3100 (100ng/ml), while the lower chamber was replaced with 20% FBS-containing medium and incubation continued. After an additional 48 h of incubation, the filters were removed, processed and the number of invaded cells counted as previously described [34 (link)–36 (link)]
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Cell Invasion Assay with HuR-FNP Treatment
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Other organizations : University of Oklahoma Health Sciences Center
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Variable analysis
independent variables
- HuR-FNP treatment
- Number of invaded cells
- Seeding of 5 × 10^4 cells in the upper chambers
- Serum-free medium in the lower chambers
- 6 h of transfection
- Replacement of culture medium in the upper chamber with 2% serum-containing medium
- Addition of AMD3100 (100ng/ml) to the 2% serum-containing medium in the upper chamber (optional)
- 20% FBS-containing medium in the lower chamber
- Additional 48 h of incubation
- Not specified
- Not specified
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