Murine Spleen Cell Immunophenotyping

The spleens were harvested from the mice (n = 4 per genotype). Cells were released by pressing the spleens under sterile metal screens and erythrocytes were lysed in cold ammonium chloride-based buffer. Fc receptors were blocked with TruStain FcX anti-mouse CD16/CD32 (cat# 101320; BioLegend, San Diego, CA), and surface antigens were stained with fluorescence-labeled antibodies to mouse CD3, CD4, CD8, B220/CD45, IgM, CD21/35, CD23, CD19, and CD138 (Additional file 2). Data acquisition and analysis were performed as before [28 (link)] using a FACSCanto II instrument and BD FACSDiva software (version 5.0).

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Markovics A., Toth D.M., Glant T.T, & Mikecz K. (2020). Regulation of autoimmune arthritis by the SHP-1 tyrosine phosphatase. Arthritis Research & Therapy, 22, 160.

Publication 2020

TruStain FcX anti-mouse CD16/CD32 FACSCanto II instrument BD FACSDiva software

Ammonium chloride Buffer Cd138 Cells Cold Erythrocytes Fc receptors Fluorescence antibodies Genotype Metal Mouse Sterile Surface antigens

Corresponding Organization :

Other organizations : Rush University Medical Center

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Variable analysis

independent variables

Genotype of mice

dependent variables

Cell populations (CD3, CD4, CD8, B220/CD45, IgM, CD21/35, CD23, CD19, CD138)

control variables

Number of mice per genotype (n = 4)
Sterile metal screens used to release cells from spleens
Ammonium chloride-based buffer used to lyse erythrocytes
Fc receptors blocked with TruStain FcX anti-mouse CD16/CD32
Fluorescence-labeled antibodies used to stain surface antigens
Data acquisition and analysis performed using FACSCanto II instrument and BD FACSDiva software (version 5.0)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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