Transwell migration and invasion assays

Yawei Wang; Yingying Sun; Chao Shang; Lili Chen; Hongyu Chen; Dake Wang; Xianlu Zeng

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Transwell migration and invasion assays

YW Yawei Wang

YS Yingying Sun

CS Chao Shang

LC Lili Chen

HC Hongyu Chen

DW Dake Wang

XZ Xianlu Zeng

This method is extracted from research article: Cell Death Dis, Feb 2021

Distinct Ring1b complexes defined by DEAD-box helicases and EMT transcription factors synergistically enhance E-cadherin silencing in breast cancer

DOI: 10.1038/s41419-021-03491-4

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Cells were collected and resuspended in FBS-free DMEM. Twenty thousand cells were placed on 8-μm pore transwell filters (Corning, USA). For invasion assays, filters were coated with Matrigel (BD, UAS) in advance and for migration assays, this step was omitted. DMEM with 10% FBS was added to the bottom chamber as an attractant. Following incubation for 24–36 h, nonmigrated cells at the top of the filter were removed and cells at the bottom of the filters were fixed with 4% paraformaldehyde and stained with crystal violet. The total number of invaded cells in each chamber were quantified by counting at least four randomly chosen fields under ×20 magnification using a bright field microscope (IX71; Olympus).

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