The number of viable cells seeded onto a 96-well culture plate was quantified using Premix WST-1 Cell Proliferation Assay System (TaKaRa, Kyoto, Japan) according to the manufacturer’s instructions. Briefly, 10 μl of Premix WST-1 per 100 μl of culture medium was added to each well and the cells were incubated under the standard culture condition for 1 hour. WST reduction was determined with an automated ELISA plate reader, ImmunonMini NJ-2300 spectrophotometer (InterMed, Tokyo, Japan), at an optical density (OD) of 450–650 nm, as we described previously47 (link).
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