In nite m200 spectrophotometer

Cell proliferation was measured using the CCK-8 kit (Beyotime). Brie y, after transfection, the cells were digested and collected, washed with PBS, and then resuspended in DMEM medium to 5×10 4 cells/ml. The cell suspensions were added to a 96-well culture plate at 100 μl/well. The cells were incubated for 24, 48 and 72 h, respectively, and then 10 μl of CCK-8 solution was added to each well and incubated for 2 h.
The absorbance was determined at 450 nm using an In nite M200 spectrophotometer (TECAN). Six duplicates for each group.

Cell counting kit 8 (CCK8) was used to measure cell viability. An In nite M200 spectrophotometer (Tecan) was used to detect the spectrophotometric absorbance of each sample at 450 nm. Transwell assays were used to assess cell migration. BioCoat Matrigel Invasion Chambers (BD Biosciences) were used to assess cell invasion. Three random areas were assessed to determine the cell counts for the cell migration and invasion experiments. An annexin V-FITC apoptosis detection kit (Invitrogen) was used for annexin V and propidium iodide staining of cells, and the percentage of apoptotic cells was determined by ow cytometry (Beckman, USA). To assess the cell cycle, 48 hours after transfection, the cells were stained with propidium iodide and assessed by uorescence-activated cell sorting (FACS).