Cell counting Primary human lung fibroblasts were plated at a density of 5 × 104 cells per well in a 24 well plate. The next day cells were synchronized for 24 hours using DMEM/F12 medium supplemented with 0.1% FCS and antibiotics. Cells were stimulated with DMEM/F12 medium with 10...
Meanwhile, MDA-MB-231 cells were seeded into 6-well cell culture plates, which are to be infected by the lentivirus. The resulting supernatant from the transfected HEK293T cells after 48 h became the lentivirus, which was then harvested and filtered through a 0.45 μm filter. Polybrene...
2.3. In-vitro cell proliferation assays Mitochondrial activity, as a surrogate measure for cell viability, was measured using WST-1 assay. Cells were seeded at 3x103 cells/mL in a 96-well plate in triplicate. After 24 h the cells were treated with 0–105 nM of αvβ6-targeted SG3299 ...
For carfilzomib treatment, MM.1S and U266 cells were seeded into a 96-well plate at a density of 1 × 104 cells per well. After 24 h of initial seeding, each well was treated with carfilzomib (Sigma-Aldrich) at concentrations ranging from 0.01 to 1000 nM. Exosome uptake experiment ...
add 250 ml of the diluted cell suspension to each well of the labeled 96- well, flat bottom, tissue culture plate(s). RESUSPEND THE CELL SUSPENSION OFTEN DURING THE SEEDING PROCEDURE TO ENSURE A UNIFORM NUMBER AND DISTRIBUTION OF CELLS INTO EACH WELL BY PIPETTING UP AND DOWN A FEW TIMES ...
2.3. Cell atrophy model After seeding C2C12 cells in a 6-well plate (1 × 106 cells/well), when the cells were 100% confluent, they were differentiated into myotubes for 96 h in a differentiation medium. Muscle atrophy was induced by treatment with 50 nM of DEX (Sigma-Aldrich, St. Lou...
Cell proliferation assay To assess cell proliferation, the indicated Eca109 and TE-1 cells were seeded in 96-well plates and incubated under standard conditions in complete medium. Twenty-four hours after seeding, cells were incubated with 50 μM EdU (RiboBio, Guangzhou, China) for 6 h ...
Cell Counting Kit-8 (CCK8) assays For the CCK-8 assay, AGS, SGC-7901, and MKN-45 cells were seeded into 96-well plates at a density of 1,200 cells per well. The optical density at 450 nm (OD450) of each well was measured at 0, 24, 48, 72, and 96 h after seeding, followi...
The transfection began by seeding 400,000 cells in a six-well cell culture dish. The next day, 0.2 μg of the desired Siglec DNA in pCDNA, 2 μg of pOG44 plasmid, and 7 μg of Lipofectamine Plus reagent (Thermo Fisher) were added to 250 μL of Opti-MEM (Gibco) and ...
One million keratinocytes were transfected with control siRNA or SPT6 siRNA 1 day before seeded on a 12-well plate (Wuxi, CELL NEST, China, Cat# 712001). After inducing differentiation for 3 days, the cells were washed briefly with phosphate-buffered saline (PBS) two times. Permeabilized nucl...