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In total, 50 L of the diluted Matrigel gel was added to the upper chamber of the transwell, gently tilted to ensure even spreading, and incubated at 37 C for 4 h in a cell culture incubator

Kretzschmar, M., Liu, F., Hata, A., Doody, J

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This enzymatic action depletes the available intracellular pool of nicotinamide, directly limiting the synthesis of nicotinamide adenine dinucleotide (NAD+)

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