Animal13neck: Створена сторінка: 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, U...

2017-02-25T14:19:34Z

Створена сторінка: 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, U...

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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 [http://www.selleckchem.com/products/NVP-AUY922.html 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 [http://www.selleckchem.com/products/RO4929097.html 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 [https://en.wikipedia.org/wiki/Fleroxacin 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

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Animal13neck: Створена сторінка: 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, U...