Cell count kit 8 assay

Cell viability was determined by Cell Count Kit-8 assay (CCK-8; Beyotime, China). Cells were seeded in 96-well plates at a density of 1 × 10⁴ cells per well in 100 µl complete medium. For H9C2 cells, different gradient concentrations of LPS (1, 2.5, 5, 7.5, 10 µg/ml) were added, while AC16 cells were exposed to 10 mg/ml LPS. The cells were then incubated for 24 h. After that, 10 µl CCK-8 solution was added to each well and incubated for another 2 h. Finally, the optical density (OD) value of each well was measured at 450nm using the microplate reader (PerkinElmer, Turku, Finland). Cell activity histograms were drawn according to OD value. Statistical analysis was conducted using independent sample t-tests, and P < 0.05 was considered statistically significant.

The cell viability was determined using the Cell Count Kit-8 assay (CCK-8) (Beyotime Institute of Biotechnology, Shanghai, China). The effect of the aqueous extract of C. monnieri (L.) on the activity of T. rubrum was detected by the CCK-8. The experimental strains were divided into four groups. The dry powder of C. monnieri (L.) was adjusted to 64, 128, and 256 μg/ml concentrations, and C. monnieri (L.) aqueous extract was added using RPMI-1640 medium; and these were used as the three experimental groups. The fourth group was the control group and did not add any aqueous extract. In a 96-well cell culture plate, 100 μl of fungal suspension was added at the corresponding position, and 100 μl of the three groups with different C. monnieri aqueous extract concentrations was added. The control group was treated with 100 μl of fungal suspension and 100 μl of RPMI-1640 medium. Each sample was repeated three times. After incubation at 28°C for 0.5, 12, and 24 h, the samples were treated in strict accordance with the operation instructions of CCK-8 kit. Ten microliters of CCK-8 solution was added to each well and incubated for another 2 h to measure the cell viability. The optical density of the CCK-8 solution was measured at 450 nm in accordance with the kit instructions.

Cell count KIT-8 assay (C0038, Beyotime) was utilized to detect the proliferation capacity of normal and ADAMS12-deficient GC cells. 2 × 10³ cells were plated in 96-well plates with the 450 nm absorption being recorded every 24 h.
For Transwell migration and Matrigel invasion, after 24 h incubation, the migrated cells were fixed with methanol and stained with crystal violet. The migrated and invaded cells were captured and calculated using microscope.