To optimize the assay, we used HepG2 and HK-2 cells at confluency 50–70%. After treatment with tested compounds (CisPt, CAM, STA, NPs) for 6, 24 and 48 h, the cells grown in a 96-well plate were centrifuged (5 min, 8000g) at RT. Then, 70 µL of a supernatant was ...
20 nM of siRNAs were applied to transfect rPT cells (40% confluency) with RNAiMAX Transfection Reagent following the manufacturer’s specification. The medium was refreshed in DMEM/F12 medium after 5 h at 37 °C. As indicated, cells were treated with 2.5 µM Cd and/or 1 µg/ml Qu ...
Cells cultured to 90% confluency were detached then seeded in a single well (1 × 105cells) of a poly-lysine coated, clear bottom, black wall 96 well plate (Greiner Bio-One). Following incubation for 24 h at 37 °C, the culture media was replaced with 100 μL DMEM contain...
(A) After microporation, cells survival was analyzed by the MTS assay at different time points under standard culture conditions (a-MEM containing 10% FBS and subculturing at 90% confluency). (B) pEGFP-N1 vector was transfected into different hUCB-MSCs from different donors by microporation....
In MDCK epithelial cells, cell contact at confluency initiates a protracted process of morphogenesis during which several proteins known to bind the cytoskeleton become progressively associated with the detergent-resistant cell fraction and distributed to their characteristic polarized domains. Using extraction...
All GEECs were plated at a density of 7.5×105 in 6-well plates and were transfected at 50% confluency using Lipofectamine 2000 (Invitrogen, Waltham, MA, USA). 2.3. Dual-luciferase reporter assay To verify the target genes of miR-26a, bioinformatics analysis of miRNA-binding sequences on ...
At differentiation days between 28 and 32, HNK1−ERBB3+NGFR+ cells were collected and cultured in skeletal muscle growth media (see Table 1) with medium change every other day. When the SMPCs reached 70–80% confluency (designated as passage 0), they were expanded and then cryopreserved....
Cell viability was determined colorimetrically by CellTiter96® cell proliferation assay, using the dehydrogenase substrate 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; MTS. Cells were plated into a 96-well plate, cultured to about 90% confluency, th...
Cells were seeded at 5.2 × 104 cells/cm2 and split 1:2 when they reached 80% confluency. However, after one to three passages, the cells cultured in USSC growth mediumACF grew as non-adherent spheres without requiring any intervention to initiate aggregation. To maintain sphere proliferation...
At 48 h after transfection with WT, S3D, S3A, shCFL1, and control vectors, 5 × 105 cells/well were seeded into six-well plates. The cells were scraped with a 200 µL pipette tip when they achieved 90% confluency. After washing with phosphate-buffered saline, the scraped cells were...