In cell proliferation assay, 1×104 cells/well were seeded into a 96-well plate. At different experimental end time point, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was carried out using the MTS assay kit (Thermofisher, Shanghai, China) according to the manufacturer’s protocol.
In 5-Bromo-2′-deoxyuridine (BrdU) assay, the cells were cultured for three days according to designed experimental condition before labeling with (3 µg/mL) BrdU (Thermofisher) for 4 h. The analysis of BrdU was performed as previously described.9 (link)
In colony formation assay, 1 × 104 cells/well were seed in the 6-well plate after designed experimental treatment. After 2 weeks, cell colonies were fixed in 10% formalin and stained with crystal violet (0.1% w/v).
The apoptosis of cell was analyzed by Hoechst-33,258 (Thermofisher) nuclear staining. After treatment, the cells were stained with Hoechst-33,258 staining medium (0.1% Hoechst, 0.5% NP-40, 4% formaldehyde in PBS). At least 400 cells in each group were count, and the cells with fragmented nuclear were considered as apoptosis cells.
In 5-Bromo-2′-deoxyuridine (BrdU) assay, the cells were cultured for three days according to designed experimental condition before labeling with (3 µg/mL) BrdU (Thermofisher) for 4 h. The analysis of BrdU was performed as previously described.9 (link)
In colony formation assay, 1 × 104 cells/well were seed in the 6-well plate after designed experimental treatment. After 2 weeks, cell colonies were fixed in 10% formalin and stained with crystal violet (0.1% w/v).
The apoptosis of cell was analyzed by Hoechst-33,258 (Thermofisher) nuclear staining. After treatment, the cells were stained with Hoechst-33,258 staining medium (0.1% Hoechst, 0.5% NP-40, 4% formaldehyde in PBS). At least 400 cells in each group were count, and the cells with fragmented nuclear were considered as apoptosis cells.