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To check the viability of the adherent neurons after the NMDA and AgmF treatment we performed crystal violet staining of the neurons. After treating the cells with NMDA followed by Agmatine and AgmF we washed the cells with PBS for 2 times and stained the cells with crystal violet (0.5% in water/methanol 4/1) for 5 min and then rinsed with water and left to dry overnight [26]. The images were taken by using a light microscopy. Cell number was counted from 4–5 images obtained from 4 independent experiments.
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