Luminespib : An Ultimate Leisure!

The resulting cells were fixed with 1% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA) prior to be analyzed using FACS caliber (BD Biosciences, San Jose, USA) equipped with a CELLQuest software. Cytotoxicity assay A549 cells (4 �� 103 cells suspended in 100 ��L of RPMI 1640 supplemented with 10% FBS) were seeded onto a 96-well plate (SPL Lifesciences, Pocheon, Korea) and incubated overnight at 37��C with 5% CO2 to allow the cells to adhere to the plate. Next day, the culture supernatant was removed, and the antibody (dissolved in serum-free RPMI at a concentration of 10 ��g/mL), either alone or in combination with 1 ��g/mL of cisplatin, was added to the cells. Cells in fresh medium without antibody or cisplatin were used as control. Plates were incubated at 37��C for a given incubation period, after which 10 ��L Luminespib clinical trial of the solution from EZ-Cytox Cell Viability Assay kit was added to each well and incubated at 37��C with 5% CO2 for 2 hours. The extent of reduction to formazan within cells was quantified by measuring absorbance at 480 nm using SpectraMax M5 (Molecular Devices, Sunnyvale, USA). Apoptosis assay A549 cells (1 �� 105) in 100 ��L of RPMI 1640 supplemented with 10% FBS were seeded onto a 6 well plate (SPL Lifesciences, Pocheon, Korea). Plates were incubated overnight at 37��C with 5% CO2 to allow cells to adhere to the plate. The culture supernatant was removed, and the antibody (dissolved in serum-free RPMI at a concentration of 10 ��g/mL), either alone or in combination with 1 ��g/mL of cisplatin, was added to cells. Cells in fresh medium without antibody or cisplatin learn more were used as control. Apoptotic cell death was assessed 48 hours later by determining the percentage of cells positive for FITC-labeled Annexin V and PI. The proportion of Annexin V/PI positive Fleroxacin cells were analyzed using FACS caliber equipped with CELLQuest software. Statistical analysis Data were expressed as mean SD. Comparisons between experimental groups were made by one-way ANOVA. The differences in mean values were considered significant at P