ANTI-SOMATOSTATIN ICDCLS PUR

Immunohistochemistry of formalin-fixed paraffin embedded human pancreas tissue using 0.1 µg/mL of Mouse IgG1 K Isotype Control Purified (left) or 0.1 µg/mL of Anti-Somatostatin Purified (right) followed by Anti-Mouse IgG Biotin, Streptavidin-HRP, and DAB visualization.Nuclei are counterstained with hematoxylin.

Figure 2 DBP Alters alpha Cell and delta Cell Number and Size (A and B) Cell resolution reconstruction of pancreatic sections reveals no differences in alpha cell and beta cell mass in DBP +/+ and DBP -/- mice (A), quantified in bar graph (B) (scale bar represents 530 mum; representative images are shown; inset is a zoom showing maintenance of cellular resolution in a single image; n = 9-12 sections from 3 to 4 animals; unpaired t test). (C-I) Morphological analyses of DBP -/- islets (C) reveal increased alpha cell number (D) and decreased alpha cell size (E) (representative images shown in right panel), but normal area occupied (F). By contrast, beta cell number is increased (G), although beta cell size (H) (representative images shown in right panel) and area (I) are unchanged (scale bar in D represents 85 mum; scale bars in E and H represent 10 mum; n = 24-45 islets from 3 to 4 animals; E and H are zooms of C to better show alpha cell and beta cell size; DAPI is shown in blue; unpaired t test). (J-M) delta cell proportion (J and K; n = 55-79 islets from 4 to 5 animals) and size (L and M; n = 29 to 30 islets from 3 animals) are decreased in DBP -/- islets (scale bar in J represents 85 mum; scale bar in L represents 10 mum; representative images are shown; L is a zoom of J to better show delta cell size; unpaired t test). (N) Expression levels of the alpha cell differentiation markers Arx , Pax6 , Pou3f4 , and Irx2 are similar in DBP +/- and DBP -/- islets (n = 3-10 animals;

Fig. 4 LUXendin645 reveals GLP1R expression in a subpopulation of alpha-cells. a - c LUXendin645 labeling is widespread throughout the intact islet, co-localizing predominantly with beta-cells a and delta-cells b , but less so with alpha-cells c stained for insulin (INS), somatostatin (SST), and glucagon (GCG), respectively ( n = 18 islets, seven animals, three separate islet preparations) (scale bar = 26 um). d Following dissociation of islets into cell clusters, LUXendin645 labeling can be more accurately quantified (arrows highlight cells selected for zoom-in) (scale bar = 26 um). e Zoom-in of d showing a LUXendin645 - (left) and LUXendin645 + (right) alpha-cell (arrows highlight non-labeled cell membrane, which is not bounded by a beta-cell) (scale bar = 26 um). f Box-and-whiskers plot showing proportion of beta-cells (INS) and alpha-cells (GCG) co-localized with LUXendin645 ( n = 18 cell clusters, ten animals, three separate islet preparations) (box and whiskers plot shows range and median; mean is shown by a plus symbol). g Ins1Cre Thor ;R26 mT/mG dual fluorophore reporter islets express tdTomato until Cre-mediated replacement with mGFP, allowing identification of beta-cells (~80% of the islet population) and non-beta-cells for live imaging (scale bar = 26 um). LUXendin645 (LUX645) highlights GLP1R expression in nearly all beta-cells but relatively few non-beta-cells ( n = 31 islets, six animals, three separate islet preparations). h A zoom