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Cultured cells were seeded in 12 wells plate at density of 80,000 cells per well and incubated.
96 wells plate were seeded with 20000 Hospicells.
Total RNA was isolated at 6 h after glucose treatment from the H9C2 cardiomyoblasts cell cultured in 6 wells plate using Trizol Reagent.
Each well of the 96 wells plate was filled with 150 µl 10% crystal violet (Harleco, Gibbstown, NJ), and incubated for 30 min at room temperature.
For actin cytoskeleton staining, 3,5.104 hcellsD3 cells were plated onto collagen-coated coverslides, in EBM2 medium plus BSA 1%, and placed in 24 wells plate.
Briefly, samples containing the same concentration of total protein were incubated for 2 h in a 96 wells plate coated with IR β subunit-specific monoclonal antibody.
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The 96-wells plate was centrifuged at 2000 rpm for 10 min to settle the suspended cells at the bottom.
One day before transfection, 120,000 cells were seeded in a 6-wells plate without antibiotics.
0.1×106 cells were transiently transfected in 24-wells plate using Lipofectamine 2000 (Invitrogen, Breda, The Netherlands) according to the manufacturer's instructions.
Each primer pair was repeated 3 times on a 96-wells plate (6 measurements per genotype) and duplicate plates were used.
PCR products were purified on a 96-wells plate (Thermo Fisher Scientific, Waltham, USA) using gel filtration with Sephadex™ G-50 (GE Healthcare, Uppsala, Sweden).
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