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3) in buffer A plus 20 μM DTPA, 0.05% BSA were then added to the wells followed by 1-h incubation at room temperature. The plate was washed with buffer A. The enhancement solution was added and the fluorescence was measured after 10-min incubation on the shaker.
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57749087937
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note
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Physicochemical properties, including FPSA, were calculated using ADME Profiler: Program for discreet compound analysis and calculation of physicochemical properties, version 1.6.0; Pharmacopeia Inc., 2004.
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2 for 24 h. The media was then decanted and 200 μL of proliferation media containing EBM-2 plus 2% FBS and 3 ng/mL FGF with the compound was added and the cells were incubated for an additional 48 h. Cells were then fixed with 2.5% glutaraldehyde for 30 min, washed and stained with 0.1% crystal violet. Absorbance at 595 nM was measured after being washed and destained using 10% acetic acid.
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57749113971
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2 for 1 h at 37 °C, followed by blocking with 2% BSA in the coating buffer for 1 h at 37 °C. HUVEC cells were then seeded to the upper chamber at 20,000 per well in 100 μL of migration buffer in the presence or absence of the compound. Migration buffer (500 μL) was added to the bottom chamber followed by incubation at 37 °C for 4 h. The non-migrated cells were removed from the upper side of the filters with a cotton ball. The migrated cells were fixed with 2.5% glutaraldehyde followed by washing and staining using 0.1% crystal violet. The stained cells were photographed and analyzed using ScanImage.
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