Cell invasiveness was evaluated using a Transwell chamber assay (Costar, USA). Chamber membranes (8μm, BD Falcon) were pre-coated with 6μl matrigel at 4C overnight, and seeded with 1×105 cells. DMEM without FBS supplement was added to the upper chamber and 600μl of DMEM (containing 10% FBS) was added to the lower chamber. Cells were incubated for 48 h with or without treatment. The cells on the top of membranes were removed, and the cells that penetrated the membrane were fixed in ethanol, followed by crystal violet staining. The number of cells on the opposite side of the membrane was counted under the microscope in four random fields of vision.
Chamber membranes
Manufactured by BD
Chamber membranes are a type of lab equipment used to facilitate the separation and filtration of various substances. These membranes are designed to selectively allow the passage of certain molecules or particles while retaining others, depending on their size, charge, or other physical properties. The core function of chamber membranes is to enable controlled and efficient separation processes in a laboratory setting.
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Lab products found in correlation
3 protocols using chamber membranes
1
Transwell Cell Invasion Assay
2
Transwell Matrigel Invasion Assay
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Cell invasiveness was evaluated using a Transwell Matrigel invasion assay (Costar). Chamber membranes (8 μm, BD Falcon) were not precoated with 6 µL Matrigel at 4 °C overnight before seeding with 2 × 104 cells. RPMI with 2% FCS supplement was added to the upper chamber, and 600 µL of RPMI containing 20% FCS was added to the lower chamber. The cells were incubated for 48 h, with or without treatment. The noninvading cells on the top of the membranes were carefully removed, and the invaded cells that penetrated the membrane were fixed in ethanol, followed by crystal violet staining. The number of invaded cells was counted under the microscope in five random fields of vision, and representative images were photographed.
3
Transwell Migration Assay for GBM Cells
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For the transwell migration assay, the cultured wild-type or shUSP6NL GBM cells were incubated for 24 h in 6-well plates (Costar, Washington, DC, USA) with RPMI and 10% FCS until they reached 95%–100% confluence. Chamber membranes (8 μm, BD Falcon) were not precoated with Matrigel before seeding with 1 × 105 cells. RPMI with 2% FCS supplement was added to the upper chamber, and 600 μL of RPMI containing 20% FCS was added to the lower chamber. The cells were incubated for 48 h, with or without treatment. The noninvading cells on the top of membranes were carefully removed, and the invaded cells that penetrated the membrane were fixed in ethanol, followed by crystal violet staining. The number of invaded cells identified under the microscope in five random fields of vision were counted, and representative images were photographed.
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