Fig. 6: Induction of α-cell-enriched genes in 2-week-old βRapKO^GFP.

a GO analysis of differentially expressed genes as identified by microarray of 2-week-old WT (n = 3) and βRapKO^GFP β-cells (n = 4) was associated with β-cell function and metabolism. b Venn diagram representation of the subsets of Raptor-regulated genes in 2-week-old purified β-cells that were enriched in α-cells and heatmap showed the differential expression of nine overlapped genes in 2-week-old WT and βRapKO^GFP β-cells (n = 3–4). c Analysis strategy to identify Raptor-dependent genes in β-cells is shown. d Heatmap of seven Raptor-dependent genes obtained from 8-week-old RNA-seq and 2-week-old microarray. e Volcano plot shows differential genes between 2-week-old WT and βRapKO^GFP β-cells. Microarray identification of Etv1 and Tspan12 as significantly upregulated α-cell-enriched genes in 2-week-old βRapKO^GFP β-cells. f INS-1 cells were transfected with SiRaptor or SiNC for 48 h. qRT-PCR confirmed Raptor-dependent genes in INS-1 cells (n = 4 independent cell experiments, for Raptor, p = 0.029; for Slc2a2, p = 0.048; for Msln, p = 0.031; for Ppp1r1a, p = 0.28; for Etv1, p = 0.004; for Tspan12, p = 0.008; for Aass, p = 0.02). g, h INS-1 cells were transfected with GFP, Etv1, or Tspan12 overexpression vector. g Luciferase reporter assay using a rat glucagon promoter reporter (n = 3 independent cell experiments, p = 0.039). h qRT-PCR analysis of glucagon expression in vector transfected INS-1 cells (n = 4 independent cell experiments, p = 0.02). Data represent means ± SEM. *p < 0.05, **p < 0.01 by two-sided Student’s t-test.