Wst 1 assay kit

WST-1 assay kit (Beyotime Biotechnology) was conducted to evaluate the proliferation rate of cells in our study. Briefly, transfected log-phase cells were inoculated into 96-well plates at a density of 1,000 cells per well as described before.22 (link) Then, cell proliferation rates were detected at 24 hrs, 48 hrs, 72 hrs, and 96 hrs using SpctraMax M5 (Molecular Devices, San Francisco, CA, USA).

Cell proliferation were measured with a WST1-assay kit (Beyotime Biotechnology) as described before.3 (link) In brief, cells transfected with siRNAs or plasmid vectors and negative controls were inoculated in 96-well plates at a density of 2,000 cells per well. Then, cell-proliferation percentages were measured at 24, 48, 72, and 96 hours using a SpectraMax M5 (Molecular Devices, San Francisco, USA).

SGC-7901 cells were plated into a 96-well plate at a density of 4×10³/well and treated with BBR for 12 and 24 h at different concentrations. Then, cells were treated with WST-1 and culture was continued for 2.5 h according to the protocol of the WST-1 Assay Kit (Beyotime Biotechnology Company). Finally, the observation density was read at 450 nm by a plate reader (BioTek Epoch, Winooski, VT, USA). The percentage of proliferation inhibition was calculated using the following formula: (1–the OD value of the BBR-treated group/the OD value of the control group) ×100%. The experiment was repeated at least three times.