Corning® Transwell® polycarbonate membrane cell culture inserts

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Seed 2 × 10^5 cells per well in 200 μL serum-free medium.	- Incubate cells for 24–48 h at 37C. - Treat cells with 0.1 % crystal violet for 30 min, followed by washing with water for 30 s to remove residual dye

Upstream tips
- Seed 2 × 10^5 cells per well in 200 μL serum-free medium.
Protocol tips
- Incubate cells for 24–48 h at 37C. - Treat cells with 0.1 % crystal violet for 30 min, followed by washing with water for 30 s to remove residual dye

Publication protocol

Migration of HCC cells was assessed using the 24-well polycarbonate membrane cell migration assay kit (#3422, Corning Incorporated Costar, Tewksbury, USA) according to the manufacturer’s instructions. Briefly, HCC cell lines were transfected with control and ARID1A shRNAs and were incubated in serum-free medium for 24 h. The cells were then transferred to the upper chamber of a Transwell plate by seeding 2 × 10 cells per well in 200 μL serum-free medium. Next, 0.5 mL of 10 % fetal bovine serum-containing medium was added to the lower chamber as a chemoattractant. Cells were incubated for 24–48 h (depending upon migration capability) at 37 °C. Non-migrating cells on the upper membrane surface were scraped off using cotton swabs. Cells that migrated to the bottom of the membrane were stained with 0.1 % crystal violet for 30 min, followed by washing with water for 30 s to remove residual dye. Four views were examined per transwell and cells/view were counted at 200× magnification. Each experiment was performed in triplicate. The invasion assay was performed in a similar fashion using BD BioCoat™ Matrigel™ Invasion Chambers (#354480, BD Biosciences), except that the upper chambers were precoated with ECMatrix™ gel.

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Manufacturer protocol

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