Fig. 4: Dual agonist engages distinct endogenous GLP1R/GIPR nanodomains.
From: Fluorescent GLP1R/GIPR dual agonist probes reveal cell targets in the pancreas and brain
a, Schematic showing single-molecule labelling and localization strategy for GLP1R (LUXendin645; LUX645), GIPR (sGIP648) and GLP1R/GIPR (daLUXendin660) (PDB 7VBI)26. b, Representative dSTORM images show different single-molecule densities and organization (30 nm) of LUX645-bound GLP1R, sGIP648-bound GIPR and daLUXendin660-bound GLP1R/GIPR across an islet cell population (nucleus is bounded by dashed line) (n = 5 islets from three mice). c,d, daLUXendin660 labels more GLP1R/GIPR clusters compared to either LUX645 or sGIP648, shown by representative images (c) and bar graph (d) (n = 5 islets from three mice) (one-way ANOVA with two-stage linear step-up procedure of Benjamini, Krieger and Yekutieli). e, Distribution and mean (inset) localization per cluster are similar across LUX645, sGIP648 and daLUXendin660 probes (n = 5 islets from three mice) (one-way ANOVA with Šídák’s multiple comparisons test). f, As in e, but for cluster density (one-way ANOVA with Šídák’s multiple comparisons test). g, Single particle tracking of daLUXendin660-labelled GLP1R/GIPR, showing min–max displacement (n = 5 cells, two independent repeats). h,i, SiR-d12 can be installed on daLUXendin (h) to create daLUXendin651-d12 (i), optimized for STED nanoscopy. j, Confocal snapshots showing CHO-K1:SNAP-GLP1R:Halo-GIPR cells labelled for GLP1R (BG-Sulfo549) and GIPR (CA-AF488) before live STED imaging of daLUXendin651-d12 labelling (n = 5 wells). Scale bars are provided on each figure. ns, non-significant. Exact P values are displayed on each graph.
