Xcellligence software 2

Cell proliferation was detected using CCK-8 assay (Beyotime Institute of Biotechnology, Shanghai, China) and Real-time Cellular Analysis (RTCA assay; ACEA Biosciences, Inc.; Agilent Technologies, Inc.), as previously described [23 (link)–25 (link)]. For CCK-8 assay, cells were transfected with the plasmids and seeded in 96-well plate at a density of 5, 000–10, 000 cells/well. Cells were cultured for 24, 48, and 72 hours, followed by incubation with 10 μl CCK-8 reagent, the absorbance values at 450 nm were determined by a Spectra Max 190 reader (Molecular Devices, LLC). For RTCA assay, 5, 000–10, 000 cells were seeded into the E-plate. The data were recorded using xCELLLigence software 2.0 (ACEA Biosciences, Inc.; Agilent Technologies, Inc.) and analyzed using GraphPad Prism 5.0 (GraphPad Software, Inc.).

The RTCA xCELLLigence system (ACEA Biosciences, Inc.; Agilent Technologies, Inc.) was usually used to measure cell morphology, proliferation and migration in vitro in a non-invasive manner, as described previously [23 (link)–25 (link)]. In brief, the lower chamber was added into complete medium, while cells were seeded into serum-free culture medium to induce migration and invasion. The data were recorded using xCELLLigence software 2.0 (ACEA Biosciences, Inc.; Agilent Technologies, Inc.) and analyzed using GraphPad Prism 5.0 (GraphPad Software, Inc.).