Gastric Tissue Collection and Processing

Stomach tissue collection was performed as described previously (10 (link)). Briefly, the stomachs were opened along the greater curvature. For histology, gastric strips from both the lesser and greater curvatures were fixed in formalin for paraffin sections. For DNA, gastric tissue was snap frozen in liquid nitrogen and extracted using the DNEasy Blood and Tissue Kit (Qiagen, Valencia, CA). For flow cytometry, gastric cells were digested using a modified version of the protocol described by Geem et al. (11 (link)), which utilizes 17.9 μg/ml Liberase TM (Cat #05401119001, Roche Diagnostics Corporation, Indianapolis, IN) instead of Type VIII collagenase. Immunohistochemistry and immunofluorescence were performed as described previously (10 (link)), using antibodies against the following targets: calprotectin (#ab22506, Abcam); CD11b-AF594 (clone M1/70, #101254, BioLegend); Cxcr2-AF488 (#FAB2164G, R&D Systems); GSII-FITC lectin (#FL-1211, Vector Labs, Burlingame); H⁺/K⁺-ATPase-β (#D032-3, Medical and Biological Laboratories, Woburn, MA); intrinsic factor (gift from David Alpers, Washington University, St. Louis, MO); E-cadherin-FITC (#612130, BD Biosciences, San Jose, CA); E-cadherin-AF647 (#560062, BD Biosciences); and 4 Hydroxynonenal antibody (#ab46545, Abcam).

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Kao K.D., Grasberger H, & El-Zaatari M. (2023). The Cxcr2+ subset of the S100a8+ gastric granylocytic myeloid-derived suppressor cell population (G-MDSC) regulates gastric pathology. Frontiers in Immunology, 14, 1147695.

Publication 2023

DNEasy Blood and Tissue Kit Liberase TM calprotectin CD11b-AF594 (clone M1/70 4 Hydroxynonenal antibody

4 hydroxynonenal Af647 Antibodies Antibody Biological Blood Calprotectin Cd11b Cells Clone Collagenase Diagnostics E cadherin Fitc Flow cytometry Formalin Frozen Gastric H k atpase Immunofluorescence Immunohistochemistry Intrinsic factor Lectin Liberase Nitrogen Paraffin Tissue Vector

Corresponding Organization : University of Michigan–Ann Arbor

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Variable analysis

independent variables

Tissue collection method (formalin fixation, snap freezing in liquid nitrogen)

dependent variables

Histology
DNA extraction
Flow cytometry
Immunohistochemistry
Immunofluorescence

control variables

Gastric tissue from both the lesser and greater curvatures
Use of antibodies against specific targets (calprotectin, CD11b, Cxcr2, GSII lectin, H+/K+-ATPase-β, intrinsic factor, E-cadherin, 4 Hydroxynonenal)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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