C28/I2 cells were seeded on 96-well plates and were stimulated with different doses of LPS (1, 5 and 10 μg/mL) for 12 h. After that, cells were incubated with 20 µL of Methyl thiazolyl tetrazolium (MTT, Thermo Fisher Scientific) solution for 4 h at 37°C. After dissolving the formazan crystals, the absorbance was measured by a Microplate Reader (Bio-Rad, Hercules, CA, USA) at 490 nm.
Apoptosis analysis was performed by an Annexin V fluorescein isothiocynate (FITC) and propidium iodide (PI) apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) using Flow cytometry. C28/I2 cells were stimulated by LPS or transfected for 48 h. Then cells were harvested and stained with 5 µL V-FITC as well as 5 µL PI for 20 min in the absence of light. Subsequently, cell apoptosis was determined by a flow cytometer (BD Biosciences).
Apoptosis analysis was performed by an Annexin V fluorescein isothiocynate (FITC) and propidium iodide (PI) apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) using Flow cytometry. C28/I2 cells were stimulated by LPS or transfected for 48 h. Then cells were harvested and stained with 5 µL V-FITC as well as 5 µL PI for 20 min in the absence of light. Subsequently, cell apoptosis was determined by a flow cytometer (BD Biosciences).