Transwell Real-Time Cell Migration Assay

Transwell migration assays were performed as previously described [17] (link), [19] (link). To measure impedance-based real-time cell migration, cells were transfected with plasmids of interest as indicated. 24 hours (overexpression) or 48 hours (shRNA) after transfection, cells were re-seeded on Transwell CIM-plate 16 plates from Roche (Indianapolis, IN). After attachment of cells (3 hours after re-seeding for HeLa, 1 hour for MDA-MB-468), cell migration towards NIH-3T3-conditioned media was continuously monitored in real-time for indicated hours) using the xCELLigence RTCA DP instrument (Roche). Error bars (grey) represent three experiments.

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Döppler H., Bastea L.I., Borges S., Spratley S.J., Pearce S.E, & Storz P. (2014). Protein Kinase D Isoforms Differentially Modulate Cofilin-Driven Directed Cell Migration. PLoS ONE, 9(5), e98090.

Publication 2014

Transwell CIM-plate 16 plates xCELLigence RTCA DP instrument

Cell migration Cells Cells attachment Conditioned media Hela Migration assays Nih 3t3 cell Plasmids Rtca Shrna Transfection

Corresponding Organization : Mayo Clinic in Florida

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Variable analysis

independent variables

Plasmids of interest (overexpression or shRNA)

dependent variables

Cell migration towards NIH-3T3-conditioned media

control variables

Time of transfection (24 hours for overexpression, 48 hours for shRNA)
Cell type (HeLa, MDA-MB-468)
Time of cell attachment (3 hours for HeLa, 1 hour for MDA-MB-468)
Transwell CIM-plate 16 plates from Roche (Indianapolis, IN)

controls

Positive control: NIH-3T3-conditioned media
Negative control: Not specified

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