Fig. 6: Overexpression of EDN3 in stem cell-derived islets mitigates hypoxia effect in vitro and in vivo.
From: Improving cellular fitness of human stem cell-derived islets under hypoxia
a Schematics of the experiment setup. b EDN3 gene expression enrichment in native human tissue cell types. c A uniform manifold approximation and projection (UMAP) visualization of control SC-islets cultured in 2% or 21% oxygen or EDN3 overexpressed SC-islets cultured in 2% oxygen for 4 weeks. d Gene expression of EDN3 in SC-β cells (Ctrl) and EDN3 overexpressed SC-β cells (EDN3+). e Percentage of high insulin gene expression in SC-β cells (Ctrl) and EDN3 overexpressed SC-β cells (EDN3+) cultured in 2% oxygen for 4 weeks in vitro. f Dot plot of pancreatic β cell gene markers and gene expression in different conditions. g Dot plot of selected upregulated genes and downregulated genes in EDN3+ SC-β cells cultured in 2% oxygen compared to SC-β cells (Ctrl) cultured in 21% and 2% oxygen. h A uniform manifold approximation and projection (UMAP) visualization of SC-β cells and EDN3 overexpressed SC-β cells transplanted in NSG mice in the subcutaneous space for 1 week. i Gene expression of EDN3 in SC-β cells (Ctrl) and EDN3 overexpressed SC-β cells (EDN3+) transplanted in NSG mice in the subcutaneous space for 1 week (n = 2 biological replicates). The whiskers extend to the minimum and maximum values. The center line indicates the median. The box boundaries represent the 25th and 75th percentiles. j Cell type enrichment analysis of SC-β cells (Ctrl) and EDN3 overexpressed SC-β cells (EDN3+) transplanted in NSG mice in the subcutaneous space for 1 week. k A volcano plot of the differential gene expression of EDN3 overexpressed SC-β cells (EDN3+) and SC-β cells (Ctrl) transplanted in NSG mice in the subcutaneous space for 1 week. The data were analyzed by a two-tailed Wilcoxon test and adjusted by the Benjamini-Hochberg method. l Static human insulin secretion of control SC-islets (Ctrl) and EDN3 overexpressed SC-islets (EDN3+) cultured in 21% and 2% oxygen for 1 week in a glucose-stimulated insulin secretion (GSIS) assay (n = 5 biological replicates). The data were expressed as means ± SD. A Two-tailed Student’s t test was performed between the two groups. One-way ANOVA followed by Tukey’s test was performed between multiple groups. Schematics of the experiment setup created in BioRender. Wang, K. (2025) https://BioRender.com/t41ycxm.
