For cell proliferation, cells at a density of 1.5 × 105/well were seeded in 12‐well plates. After treatment, cells were harvested and determined cell number using trypan blue (Solarbio, C0040) stain by cell counting plate. For MTT assay, cells were seeded in 96‐well plate with a density of 5 × 103 per well in 200 uL of complete media. After treatment, the cultures were washed with PBS (phosphate buffered solution, Solarbio, P1022) for three times. Cytotoxicity was assessed using MTT (3‐[4,5‐dimethylthiazol‐2yl]‐2,5‐diphenyltetrazolium bromide, Solarbio, M8180). MTT solutions 20 uL (5 mg mL−1 in PBS) along with 200 uL of fresh, complete media were added to each well, and plates were incubated for 4 h. After incubation, add 200 µL DMSO (Solarbio, D8371) into each well. Wrap plate in foil and shake on an orbital shaker for 5 min. Occasionally, pipetting of the liquid may be required to fully dissolve the MTT formazan. Absorbances were measured using microplate reader at 490 nm and results were expressed as % viability which was directly proportional to metabolic active cell number. Percentage (%) viability was calculated as: Cell Viability (100%) = OD in treatment well/OD in control(PBS) well × 100.
For CCK8 assay, the cell proliferation was measured using the Cell Counting Kit‐8 (NCM Biotech, C6005). The transfected cells were plated into 96‐well plates at a density of 5 × 103/well/100 µL. and then 10 µL of CCK8 solution was added to each well, followed by incubation for 2 h. The cell viability was determined by measuring the absorbance at 450 nm.
For CCK8 assay, the cell proliferation was measured using the Cell Counting Kit‐8 (NCM Biotech, C6005). The transfected cells were plated into 96‐well plates at a density of 5 × 103/well/100 µL. and then 10 µL of CCK8 solution was added to each well, followed by incubation for 2 h. The cell viability was determined by measuring the absorbance at 450 nm.
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