1.Cells are maintained at 37±1℃in a humidified atmosphere containing 5%±1% carbon dioxide. Split the cells once they grow to 70-90% confluency. 2.Typically, when splitting confluent HEK 293T cells in a 1:5 ratio, confluency is reached again after 2-3 days; 1:10 ratio, 3-4 days;...
cells exhibiting enhanced green fluorescent protein (eGFP) expression were isolated, using a FACSAria III flow cytometer, and subsequently were cultivated in complete growth media for 3 to 4 days to expand into 80% of confluency. Then, the cell suspension was subjected to serial...
Approximately 2 × 106 cells (T-75 flask with 80–90% confluency) were washed with PBS and pre-starved with methionine-free DMEM supplemented with 10% dialyzed FBS for 30 min. Then, cytosolic translation was inhibited by adding cycloheximide (PanReac, Barcelona, Spain) to 200 μg/mL. After...