Fig. 3

Identification of unique snRNA-seq gene markers in human islet endocrine cells. A snRNA-seq identification of gene biotypes referring to AnnotationHub [50]. B Identification of differentially expressed genes with a p value ~ 0 and log2-Fold change greater than 1.5 for each cell cluster and log2FC < 0.85 in the snRNA-seq data using pseudo-bulk averaged heatmap. Four top genes are presented. C Dotplot projection of newly found gene markers from the snRNA-seq data on Azimuth annotated endocrine clusters. D Dotplot projection of newly found gene markers in the snRNA-seq dataset on Azimuth annotated endocrine clusters for scRNA-seq data. E Projection of the four top newly found gene makers for endocrine cells from the snRNA-seq data on UMAP: PTPRT (α-cells), ZNF385D (β-cells), LRFN5 (δ-cells), and CNTNAP5 (γ-cells). F Projection of the four top gene makers for exocrine cells from the snRNA-seq data on UMAP: REG1A (acinar cells), CFTR (ductal cells), FLT1 (endothelial cells), COL1A1 (activated stellate cells), PRR16 (quiescent stellate cells). G Representative images of RNA fluorescence in situ hybridization performed in dispersed human islet cells using the RNA scope platform with probes targeting ZNF385D (green) intron sequences present only in pre-mRNA (top) and exon sequences present in both mature mRNA and pre-mRNA. Insulin immunofluorescence (green) was used to detect β-cells and DAPI for nuclei detection (blue)