Figure 5.

Alpha-cells contribute to CXCL10 expression in islets of new-onset T1D patients. (a) Triple immunofluorescence analysis of insulin (INS, green, panel A), glucagon (GCG, blue, panel B), and CXCL10 (red, panel C) of pancreatic islets in T1D DiViD cases. Panel E: digital zoom-in of overlapping (merge) channels, showing colocalization of CXCL10 and insulin (yellow pixels) indicated by yellow arrow and of CXCL10 and glucagon (magenta pixels) indicated by red arrow. Scale bar = 50 μm. Scale bar zoom-in = 20 μm. (b) Colocalization analysis of CXCL10 and insulin (green dots) and CXCL10 and glucagon (blue dots) in pancreatic islets of T1D DiViD cases. A total of n = 50 ICIs from 6 DiViD cases were analyzed for both CXCL10-insulin and CXCL10-glucagon colocalization rate. Values are reported as the percentage of overlapping CXCL10-insulin or CXCL10-glucagon pixels over total insulin or glucagon positive pixels, according to Mander's Coefficient calculation. Exact p-value was calculated using Wilcoxon matched-pairs signed rank test. (c) Colocalization plots of CXCL10-insulin (left) and CXCL10-glucagon (right) of a recent-onset diabetic DiViD individual ICI (Case-1). Positive pixels for CXCL10 (red), insulin (green), and glucagon (blue), alongside with colocalizing pixels (CXCL10-insulin: yellow; CXCL10-glucagon: magenta), are reported in the plots. Significant colocalizing pixels are within the area delimited by white lines, representing background and threshold levels relative to each channel. Each pixel is reported as a gray-scale RGB intensity value (0–255).