Mtt cell proliferation viability assay kit

Cell proliferation was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation/viability assay kit (Sigma-Aldrich, USA). After 48 h of transfection, the cells were seeded at 1×10⁵ cells/well into 96-well plates with 6 replicate wells per group. After further 48 h of incubation, 20 μL of MTT (5 g/L) was added to each well and incubated for 4 h. After discarding the MTT, 150 μL DMSO was added to shake and fully dissolved. The absorbance OD was measured at 490 nm by a microplate reader (Thermo, USA).
The transfected cells were seeded into the culture plate at 200 cells/well, and after 2 weeks of incubation, the cells were washed with PBS and stained with Giemsa solution. The number of colonies containing >50 cells were observed under the microscope.

Cell viability assay was performed using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) Cell Proliferation/Viability Assay kit (Sigma, Germany) according to the guidelines. The absorbance (570 nm) of each well was then measured and recorded. Cell viability was calculated with the following equation: (OD_sample / OD_control) × 100.
Apoptosis rate was evaluated using an Annexin-V-Fluos and Propidium Iodide (PI) Apoptosis Detection Kit (Sigma, Germany) by fluorescence activated cell sorter (FACS) according to the manufacturer's protocol. Annexin V positive and PI negative stained (Annexin V⁺/PI⁻) cells were recorded to be at the early apoptotic stage, whereas Annexin V and PI positive stained (Annexin V⁺/PI⁺) cells were recorded to be at the late apoptotic stage. The experiments are repeated three times.

The cells were seeded in 96-well plates for cell proliferation assay using a 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltet-razolium bromide (MTT) Cell Proliferation/Viability Assay Kit (Sigma-Aldrich Chemie Gmbh, Munich, Germany) in accordance with the guidelines. The cell apoptosis was measured using an Annexin-V-Fluos and Propidium Iodide (PI) Apoptosis Detection Kit (Sigma) by a flow cytometer (Becton Dickinson, San Jose, CA, USA) according to the manufacturer’s instructions.