Cell Migration and Invasion Assay

For migration studies, a standard assay was used to determine the number of cells that traversed a porous polycarbonate membrane in response to a chemo-attractant (higher serum concentration)^{51 (link),52 (link)} using the Cytoselect 24-well cell migration assay (Cell Biolabs). Invasion was measured using BD BioCoat Matrigel invasion chambers (BD Bioscience). In both assays, cells were transfected with AMOs, and 1.5 × 10⁵ and 2.5 × 10⁵ cells per well were seeded in an upper chamber in serum free media for the migration and invasion assay, respectively. The lower chamber was filled with media containing 10% FBS. After 24 h, cells passing through polycarbonate membrane were stained and counted according to the manufacturer’s instructions.

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Oh J.M., Venters C.C., Di C., Pinto A.M., Wan L., Younis I., Cai Z., Arai C., So B.R., Duan J, & Dreyfuss G. (2020). U1 snRNP regulates cancer cell migration and invasion in vitro. Nature Communications, 11, 1.

Publication 2020

Cytoselect 24-well cell migration assay BD BioCoat Matrigel invasion chambers

Assay Cell migration assay Cells Matrigel Membrane Polycarbonate Serum Serum free media

Corresponding Organization :

Other organizations : Howard Hughes Medical Institute, University of Pennsylvania

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Variable analysis

independent variables

Transfection of AMOs (Antisense Morpholino Oligonucleotides)

dependent variables

Number of cells that traversed the porous polycarbonate membrane in response to a chemo-attractant (higher serum concentration)
Invasion measured using BD BioCoat Matrigel invasion chambers

control variables

Cell seeding density (1.5 × 10^5 cells per well for migration assay, 2.5 × 10^5 cells per well for invasion assay)
Serum-free media in the upper chamber
Media containing 10% FBS in the lower chamber
Incubation time of 24 hours

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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