Fig. 4

In vivo lineage tracing of transplanted GFP⁺BMSC in recipient mice pancreas. a Schematic representation of the experimental model to lineage trace GFP + BMSC contributing to new ß-cell generation. b Representative image from Activin-a treated GFP + BMSC recipient mouse pancreas showing GFP-labeled ß-cells by co-immunostained for GFP (green) with insulin (red). Nuclei were stained with Dapi (blue). The graph displays quantification of c total GFP+ cells and d GFP+ ß-cells within the islets of regenerating the pancreas. Data represent mean ± SEM with N = 3 mice per group. All statistical analyses were performed using Graphpad Prism software using two-way ANOVA and Bonferroni test for p value calculations. e Representative confocal microscopic images of immunostaining for insulin (red) and GFP (green) representing infiltration of transplanted GFP + BMSC in mature islets and ductal regions. Small clusters of GFP+ ß-cell present evidence for de novo BMSC-derived ß-cell formation from the transplanted BMSC. Nuclei were stained with Dapi (blue). f Proteomic characterization by western blotting and densitometric quantification of key pancreatic endocrine differentiation transcription factors from FACS sorted islet cells of 3 pooled representative mice pancreas, depicting evidence of new ß-cell differentiation markers. Chemiluminescence signals were exposed for 1–2 min and images were captured on the Gel Documentation system (GE Healthcare) and analyzed with ImageJ software. A single cropped area of key proteins from each condition is represented, while the graph represents densitometry quantification for each protein with standard deviation from 3 pooled mice cell extracts in duplicates