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Glucose Stimulated Insulin Secretion (GSIS)
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INS-1E cells were cultivated overnight in serum-free medium ± PA 400 μM ± S1P, washed with Hanks’ Balanced Salt Solution (HBSS) without Ca+2 and Mg+2 and maintained in HBSS without glucose for 2 h. Then, the cells were stimulated with HBSS with glucose 16 mM for 30 min and the supernatant later collected. The amount of insulin secreted was quantified using a Rat Insulin ELISA (Mercodia) following manufacturer’s instructions.
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