14-3-3 protein constrains insulin secretion in pancreatic beta-cells by regulating mitochondrial function

Members of the 14-3-3 protein family are often assumed to have redundant, over-lapping roles due to their high sequence homology and ubiquitous expression. To better understand the role of 14-3-3 in beta-cells, we generated beta-cell-specific 14-3-3 knockout (14-3-3KO) mice, 14-3-3KO mice displayed potentiated insulin secretion due to enhanced mitochondrial function and ATP synthesis. Overall design: Islets isolated from WT and beta14-3-3 KO mice at 12-14 weeks were dispersed after an overnight recovery. Dead cells were removed by using a dead cell removal kit and MS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany). Single-cell RNA-seq libraries were made with the 10x Genomics Chromium Next GEM Single Cell 3' Library Kit (v3.1) and sequenced using an Illumina NovaSeq6000 (100bp x2, 50k reads/cell). Approximately 9600 cells were loaded per sample with an anticipated recovery of 6000 cells.