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High Throughput NPY-Venus and Serotonin Secretion Assays for Regulated Exocytosis in Neuroendocrine Cells

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BIO-PROTOCOL
卷 8, 期 1, 页码 -

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BIO-PROTOCOL
DOI: 10.21769/BioProtoc.2680

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NPY; Serotonin; Exocytosis; Dense core vesicle (DCV); Neuroendocrine; High throughput screen; BON cells

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  1. National Institutes of Health [DK040428, DK025861, GM119158]

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Here we describe two assays to measure dense core vesicle (DCV) exocytosis-mediated cargo secretion in neuroendocrine cells. To conduct siRNA screens for novel genes in regulated DCV exocytosis, we developed a plate reader-based secretion assay using DCV cargo, NPY-Venus, and an orthogonal H-3-serotonin secretion assay. The NPY-Venus secretion assay was successfully used for a high throughput siRNA screen, and the serotonin secretion assay was used to validate hits identified from the screen (Sorensen, 2017; Zhang et al., 2017).

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