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For analysis of cells embedded in soft agar, cells (10 5 ) were suspended in 1 mL of 0.33% noble agar, seeded in the bottom of 15-mL polypropylene tubes, cooled for 5 min on ice, and then kept at 37 C in a humidified incubator in the presence of 5% CO 2 for 24 h72 h
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4 A common feature of the pathology involves an excavation of the optic disk (cup-to-disk), which is extended to its margins
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Finally, ratio of emission intensities of the donor and acceptor fluorescent proteins (FRET ratio) reports dynamics of level of the target metabolite
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