Glucose-stimulated insulin secretion assay was performed as previously described, with minor modifications (50). Briefly, INS 832/13 cells were washed and incubated with prewarmed Krebs-Ringer bicarbonate Hepes [KRBH; 0.5% bovine serum albumin, 129 mM NaCl, 5 mM NaHCO3, 4.8 mM KCl, 1.2 mM KH2PO2, 2.5 mM CaCl2, 1.2 mM MgSO4, and 10 mM Hepes (pH 7.4)] at 37°C for 2 hours. The supernatants were collected, and the cells were stimulated with KRBH containing either 2.8 or 25.5 mM glucose for 2 hours. After supernatants were collected, cells were lysed with acid ethanol, and insulin levels in supernatants and cell lysates were measured using STELLUX Chemi Human Total Proinsulin ELISA kits (ALPCO no. 80-PINHUT-CH01).

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