Glass botttom cell culture dishes

A CCK8 kit (Vazyme Biotech, China) and a Cell-Light EdU Apollo567 In Vitro Kit (RiboBio, Guangzhou, China) were used for the evaluation of cell proliferation. For CCK8 assay, cells (2 × 10³) were seeded into 96-well plates, cultured for 12, 24, 36 and 48 hours, and CCK8 reagent was then added to each well and incubated for 2 h at 37°C. The measurement of absorption was performed using microplate reader at 450 nm (ELX-800; Bio-Tek, Winooski, VT, USA). For 5-ethynyl-2′-deoxyuridine (EdU) assay, cells (2 × 10⁵) were seeded into Glass Botttom Cell Culture Dishes (Nest Biotechnology, NJ, USA), then cells were treated according to the given instruction. Prepared samples were detected with a laser confocal scanning microscopy.

Cell proliferation was evaluated using a CCK8 kit (Vazyme Biotech) and a Cell-Light EdU Apollo567 In Vitro Kit (RiboBio, Guangzhou, China). For CCK8 detection, transfected cells (2 × 10³) were seeded into 96-well plates, cultured for 24, 48, 72 and 96 h, and then CCK8 reagent was added to each well and incubated for 2 h at 37 °C. Absorption was measured by microplate reader at 450 nm (ELX-800; Bio-Tek, Winooski, VT, USA). For 5-ethynyl-2′-deoxyuridine (EdU) assay, transfected cells (2 × 10⁵) were seeded into Glass Botttom Cell Culture Dishes (Nest Biotechnology, NJ, USA), then cells were treated according manufacture instruction, and finally cell samples were detected with a laser confocal scanning microscopy. The invasion assays were assessed using the Transwell units (Corning Costar, Tewksbury, MA, USA) percolated with Matrigel (BD Biosciences). Cells (2 × 10⁴ cells/well) were seeded at the upper compartment of the chamber in DMEM without FBS, and the lower chamber was filled with DMEM including 10% FBS as chemotaxin. After incubation for 48 h, the filters were collected, fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. Non-invading tumor cells on the top of the filters were removed with cotton swabs. Cells passing through the filter were counted under a light microscopy.