=========== HEADER ===========
Readme.txt for Confocal study of G protein-coupled receptors 17 (GPR17), Chromogranin A (CHGA), Glucagon-like peptide-1 (GLP-1) and Glucose-dependent insulinotropic polypeptide (GIP) in human intestine
Item Handle: http://hdl.handle.net/11243/36
Documentation written on 2021-02-01
By Shijun Yan
Updated 2021-02-01, by Heather Coates

=========== SUGGESTED DATA CITATION ===========
Shijun, Y., Chandrashekhar A. K., Hongxia, R. (2021). Dataset for Confocal study of G protein-coupled receptors 17 (GPR17), Chromogranin A (CHGA), Glucagon-like peptide-1 (GLP-1) and Glucose-dependent insulinotropic polypeptide (GIP) in human intestine. IUPUI University Library. http://dx.doi.org/10.7912/D2/22


=========== PRIMARY STUDY INFORMATION ===========
ACKNOWLEDGEMENTS
Project title: Confocal study of G protein-coupled receptors 17 (GPR17), Chromogranin A (CHGA), Glucagon-like peptide-1 (GLP-1) and Glucose-dependent insulinotropic polypeptide (GIP) in human intestine
Funding agency: National Institutes of Health
Award Number: R01DK120772, R00DK098294, P30DK097512, T32DK064466, UL1TR002529


Investigator Name: Shijun Yan
Investigator Institution: Herman B. Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine
Investigator Address: 635 Barnhill Dr., MS2043, Indianapolis, IN 46202
Investigator Email:yanshij@iu.edu
Investigator Role (related to this dataset): data collection, data processing/cleaning, data analysis, data visualization
Investigator ID (ORCID): N/A

Investigator Name: Chandrashekhar A. Kubal
Investigator Institution: Department of Surgery, Indiana University School of Medicine
Investigator Address: Indianapolis, IN 46202
Investigator Email:renh@iu.edu
Investigator Role (related to this dataset): sample preparation
Investigator ID (ORCID): N/A

Investigator Name: Hongxia Ren
Investigator Institution: Herman B. Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine
Investigator Address: 635 Barnhill Dr., MS2049, Indianapolis, IN 46202
Investigator Email:renh@iu.edu
Investigator Role (related to this dataset): PI, data review
Investigator ID (ORCID): N/A


DATASET Description
Immunohistochemistry (IHC) staining of GPR17, CHGA, GIP and GLP1 in human gut tissue and HEK-293 cells transfected with HA-tagged human GPR17 gene. Images were acquired and analyzed using LSM-700 confocal microscope and its software ZEN (ZEISS, Oberkochen, Germany) and Image J (NIH). All images are CZI formats and require ZEN software (ZEISS) to open.

METHODS
Human gut samples were obtained from two cadavers at IU Health Methodist Hospital. They are male and aged at 26 and 30 years old. To determine the cell types of GPR17 expressing cell in human gut, we performed immunohistochemistry (IHC) staining by using antibodies for GPR17 (NLS4229; Novus, Centennial, CO), Chromogranin A (general endocrine cell marker; SC-393941; Santa Cruz Biotechnology, Dallas, Texas), GIP (marker for K cells; ab30679; abcam, Cambridge, MA) and GLP-1 (marker for L cells; NBP1-50697; Novus). Section slides were imaged and analyzed using LSM-700 confocal microscope and its software ZEN (ZEISS, Oberkochen, Germany).

To validate the specificity of GPR17 antibody, we stained GPR17 (NLS4229; Novus) and HA (#901513; Biolegend, San Diego, CA) in HEK293 cells transiently transfected by pcDNA3.1-HA-hGPR17 plasmids. HEK293 cells were transfected with pcDNA3.1 or pcDNA3.1-HA-hGPR17 plasmids for overnight. Chamber slides were imaged using LSM-700 confocal microscope (ZEISS). 10-22 randomly selected cells per image and total 10 images were quantified for staining intensity by using Image J (NIH).  


DATE(S) of DATA COLLECTION
2018-04 through 2021-01

GEOGRAPHIC LOCATION(S) of DATA COLLECTION
Indianapolis, IN, 46202, USA

DIRECTORY/FILE NAMING CONVENTIONS
Folder: IHC images for CHGA and GPR17 in human intestine (contains 42 .czi files)
Folder: IHC images for GIP and GPR17 in human intestine (contains 11 .lsm files)
Folder: IHC images for GLP-1 and GPR17 in human intestine (contains 62 .czi files)
Folder: IHC images for HA and GPR17 in HEK293 cells (contains 12 .czi files)
File: Quantification data for CHGA, GLP-1 and GPR17.xslx
File: Quantification data for HA and GPR17.xslx


FILE INFORMATION
.czi (Zeiss - proprietary file format) 
.lsm (Zeiss - proprietary file format) 


=========== CREDITS ===========
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