Fig. 5

scRNAseq analysis reveals effects of hyperinsulinemia on cell type-specific gene expression. A Schematic describing the single-cell transcriptomics experimental design and analysis. Six mouse pancreata from each genotype were dissociated to single cells, and the samples were sorted for hoechst⁺ and PI^- live cells. The live single cells were sequenced and clustered in uniform manifold approximation and projection (UMAP) space. B Unsupervised clustering of cells from 6 PK-Ins1^-/-;Ins2^+/+ and 6 PK-Ins1^-/-;Ins2^+/- mice pancreata, represented as an UMAP plot. C Numbers of cells from PK-Ins1^-/-;Ins2^+/+ (orange) and PK-Ins1^-/-;Ins2^+/- (green) mice for each cell type. The asterisk indicates a significant difference in the number of NK cells between the genotypes. D Dot plot showing selected cell type-specific markers for identifying the cell type for each cluster. The size of dots represents the fraction of cells expressing the markers. The PK-Ins1^-/-;Ins2^+/+ mouse data are shown in blue and PK-Ins1^-/-;Ins2^+/- mouse data are shown in red. The intensity of color indicates the average expression of marker genes for each cell type. E Single-cell expression of immune checkpoint receptors and ligands in each identified cell population in both genotypes. F Immunohistochemistry of Cd20 (B cell marker) for PK-Ins1^-/-;Ins2^+/+ and PK-Ins1^-/-;Ins2^+/- pancreata. Scale bars: 0.1 mm (up) and 0.2 mm (down). G–H The quantification of Cd20⁺ area for PK-Ins1^-/-;Ins2^+/+ and PK-Ins1^-/-;Ins2^+/- male (G) and female (H) mice