Cells were cultured in serum-free DMEM for 24 h before seeding at 1 × 103 cells per well in 96-well plates in control media, WT-M2 conditioned media, orTRIM59-/--M2 conditioned media. Cell proliferation was measured using Cell Count Kit-8 (CCK-8) reagent (10 μl was added to each well at the endpoint) at the indicated times according to the manufacturer’s instructions (Dojindo Laboratories, Kumamoto, Japan). In wound healing assays, melanoma cancer cells were seeded into 6-well plates and scraped using a sterile pipette tip. Images were obtained using an inverted microscope at 0 and 24 h and then analyzed by ImageJ software (NIH, Bethesda, MD, USA). For cell invasion assays, melanoma cancer cells were seeded into Matrigel-coated upper chambers of Transwell inserts (Corning, NY, USA). After 24 h of incubation, non-invading cells were scraped off with a cotton swab, and the cells on the bottom of the chamber were fixed, stained, and counted.
Matrigel coated upper chambers of transwell inserts
Manufactured by Corning
Sourced in United States
Matrigel-coated upper chambers of Transwell inserts are a laboratory product designed for cell migration and invasion assays. The Matrigel coating on the upper chambers provides a reconstituted basement membrane matrix, which serves as a barrier for cells to migrate through. This product allows researchers to study the ability of cells to penetrate and move through this extracellular matrix-like material.
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Lab products found in correlation
2 protocols using matrigel coated upper chambers of transwell inserts
1
Assessing Melanoma Cell Proliferation and Migration
2
Transwell Assay for Migration of GBC Cells
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Migration of GBC cells was characterized using a transwell assay. MiRNA-transfected TYGBK-8 and G-415 cells were treated with mitomycin (20 µg/mL; Thermo Fisher Scientific, USA) for 20 min to stop proliferation. They were then suspended and re-plated in the Matrigel-coated upper chambers of transwell inserts (Corning, USA) in 12-well plates. The upper chambers were filled with culture medium but without FCS. The lower chambers were filled with normal culture medium (with 10% FCS). Twenty-four hours later, upper chambers were removed. The lower chambers were fixed, immunostained with crystal violet, and imaged using an Olympus inverted microscope (IX71; Olympus, Japan). The number of migrated TYGBK-8 or G-415 cells were tallied for each chamber, and normalized to the cell number in control chambers.
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