Fig 1: Immunohistochemical characterization of intensely NOS1-positive neurons.(A) A large multipolar neuron in stratum pyramidale is strongly SST and NPY positive in the somatic Golgi apparatus and weakly positive for CHRM2 in the somatodendritic plasma membrane (maximum intensity projection, z stack, height 11 μm; inset, maximum intensity projection of three optical slices, z stack height 2 μm). A smaller, more weakly NOS1-positive cell (double arrow) in the lower left is immunonegative for the other molecules; a second NPY-positive cell (arrow) adjoining the NOS1+ neuron is immunonegative for the other three molecules. (B) A NOS1-positive cell and another NOS1-immunonegative cell (asterisk) at the border of stratum radiatum and lacunosum-moleculare are both positive for GRM1 in the plasma membrane and PENK in the Golgi apparatus and in granules, but only the NOS1+ cell is immunopositive for CHRM2 (maximum intensity projection, z stack, height 10 μm). (C) An intensely NOS1-positive cell in stratum radiatum is also positive for PCP4 in the cytoplasm and nucleus and for PENK in the Golgi apparatus and in granules (maximum intensity projection, z stack, height 15 μm).
Fig 2: Confirmation of predicted colocalization of NPY and pro-CCK.(A) Interneurons at the sr/slm border immunopositive for both NPY and pro-CCK (cells 1 and 2), one of which (cell 1) is also immunopositive for CALB1. A third neuron is positive only for pro-CCK and CALB1 (cell 3). (B) Interneurons at the sr/slm border immunopositive for NPY (cells 1–3), pro-CCK (cells 2 and 4), and SLC17A8 (VGLUT3, cell 2). Note SLC17A8-positive terminals targeting unlabeled cells (arrows). (A, B) Both NPY and pro-CCK are detected in the Golgi apparatus and endoplasmic reticulum surrounding cell nuclei; in addition, some axons are also immunopositive for NPY (see arrow in (A); average intensity projections, z stacks, height 6.3 µm and 10.4 µm, respectively). (C) Combined double in situ hybridization and immunohistochemistry shows that nearly all Slc17a8-expressing cells also express Npy and are immunopositive for pro-CCK (arrows), but some Npy/pro-CCK cells do not express Slc17a8 (arrowheads). Scale bars: 10 µm (A, B), 50 µm (C). sr/slm, stratum radiatum and stratum lacunosum-moleculare.
Fig 3: Immunohistochemistry showing difference in expression of neuropeptide Y (NPY) and its receptors between lesional and normal skin. NPY, NPY1R and NPY2R were all expressed in epidermis and dermis of lesional (E~G), and normal skin (A~C), with higher degree of expression when compared to normal skin. NPY was expressed in spinous layer, NPY1R in whole layer of epidermis, and NPY2R in basal layer. (D), (H) Isotype control. Scale bar=200 µm.
Fig 4: Immunohistochemistry comparison of strain c‐Fos and NPY cell population. Colocalization of neuropeptide and activity: color image (purple) c‐Fos—Fos‐like early gene expression (green); NPY—neuropeptide Y and c‐Fos in brain slices from the ARC—arcuate nucleus of the hypothalamus in SNO (n = 5)—representative of Snord116del mice and in WT (n = 5)—wild‐type control, ingesting chronic treatment 100CFE—Caralluma fimbriata extract, at 100 mg/kg/d or PLAC—placebo 200 mg maltodextrin and 50 mg cabbage leaf, with food intake stimulant, signaling reagents, 2DG—2‐deoxyglucose, induced by i.p. injection (400 mg/kg, 10 mg =25 g mouse), in comparison with the control SAL—i.p. injection of isotonic saline
Fig 5: Analysis of Cxcl14 co-expression patterns confirms predicted properties Cck.Cxcl14 cells.(A) Cxcl14-expressing cells are CGE derived: in situ hybridization for Cxcl14 combined with immunohistochemistry for YFP in the Lhx6-Cre/R26R-YFP mouse yields no double labeling. (B) Double in situ hybridization for Cxcl14 and Reln marks a population of neurons located primarily at the sr/slm border. Note Reln expression without Cxcl14 in so and slm, likely reflecting O-LM and neurogliaform cells. (C–E) Subsets of the Cxcl14-positive neurons are positive for pro-CCK or CALB1 (in situ hybridization plus immunohistochemistry), or Npy (double in situ hybridization). (F, G) No overlap was seen of Cxcl14 with Nos1 or Kit. In all panels, arrowheads indicate double-expressing neurons. Scale bars: 200 μm (A), 100 μm (B–G). b, sr/slm border region; O-LM, oriens/lacunosum-moleculare; slm, stratum lacunosum-moleculare; sm, stratum moleculare of the dentate gyrus; so, stratum oriens; sp, stratum pyramidale; sr, stratum radiatum; sr/slm, stratum radiatum and stratum lacunosum-moleculare; YFP, yellow fluorescent protein.
Supplier Page from Abcam for Anti-Neuropeptide Y antibody [8]