Journal: Bio-protocol

Article Title: Flow Cytometry Analysis of Microglial Phenotypes in the Murine Brain During Aging and Disease

doi: 10.21769/BioProtoc.5018

Figure Lengend Snippet: Antibody list for microglial phenotypes

Article Snippet: Disease-associated microglia (DAM) , CLEC7A , 9 μg/300 μL , APC , REA154 , Human , Miltenyi , 130102985 , 1:50.

Techniques:

Journal: Cell Death & Disease

Article Title: Expansion of monocytic myeloid-derived suppressor cells ameliorated intestinal inflammatory response by radiation through SOCS3 expression

doi: 10.1038/s41419-021-04103-x

Figure Lengend Snippet: Mice were exposed to 7.5 Gy for 1 week to induce intestinal inflammation. A Photomicrographs of colon sections, histological grades, colon lengths, body weights, and survival rates for mice 21 days after irradiation. Scale bar = 100 μm B Representative dot plots of the gating strategy used to identify CD11b + /GR1 + cells from the spleens and intestines of mice 21 days after irradiation. C Representative immunofluorescence images of spleen and intestine stained for CD11b (green), GR1 (red), and CD11b + /GR1 + (yellow) cells. DAPI staining (blue) was used to determine the number of nuclei. Scale bar = 100 μm D CFSE-labeled CD3 + T cells were co‑cultured with splenic MDSCs and stimulated using anti-CD3 and anti-CD28 monoclonal antibodies. The cells were harvested and analyzed after 7 days. CFSE dilution demonstrated that T-cell proliferation was inhibited by MDSCs (left) and that this inhibition occurred in a dose-dependent manner (right). E The two MDSC subtypes, namely gMDSCs (CD11b + , Ly6G + , and Ly6C low ) and mMDSCs (CD11b + , Ly6G - , and Ly6C high ), were identified and analyzed by flow cytometry from spleen and intestine samples. The data are represented as the mean ± SD (n = 8). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. control. IR irradiation.

Article Snippet: The tissue sections were incubated with primary antibodies against CD11b (CST, 17800, 1:100 dilution, IL6 (CST, 12912, 1:100 dilution), IL10 (Abcam, ab192271, 1:100 dilution), and SOCS3 (Abcam, ab16030, 1:100 dilution), phospho-STAT3 (CST, 9145, 1:100 dilution), and GR1 (R&D Systems, MN, USA, MAB1037-100, 1:100 dilution) overnight at 4 °C.

Techniques: Irradiation, Immunofluorescence, Staining, Labeling, Bioprocessing, Inhibition, Flow Cytometry, Control

Journal: Cell Death & Disease

Article Title: Expansion of monocytic myeloid-derived suppressor cells ameliorated intestinal inflammatory response by radiation through SOCS3 expression

doi: 10.1038/s41419-021-04103-x

Figure Lengend Snippet: Mice were exposed to either 0.5 Gy or 5 Gy total body irradiation and analyzed seven days later. Representative dot plots of the gating strategies. A The population of CD11b + /GR1 + cells. B gMDSCs (CD11b + , Ly6G + , and Ly6C low ) and mMDSCs (CD11b + , Ly6G - , and Ly6C high ) from the spleens and intestines of irradiated mice were identified and analyzed by flow cytometry. C Representative dot plots of the gating strategy used to identify IL10 + cells. IL10 + cells were gated and the proportions of CD11b + /GR1 + cells. These cells were gated on gMDSCs (CD11b + , Ly6G + , and Ly6C low ) and mMDSCs (CD11b + , Ly6G − , and Ly6C high ) cells were evaluated from mouse spleens and intestines. D Representative immunofluorescence images of cells from spleen stained for GR1 (green), IL10 (red), and GR1/IL10 (yellow). DAPI staining (blue) was used to determine the number of nuclei. Scale bar = 100 μm. The data are represented as mean ± SD ( n = 8). ** P < 0.01, *** P < 0.001 vs. control.

Article Snippet: The tissue sections were incubated with primary antibodies against CD11b (CST, 17800, 1:100 dilution, IL6 (CST, 12912, 1:100 dilution), IL10 (Abcam, ab192271, 1:100 dilution), and SOCS3 (Abcam, ab16030, 1:100 dilution), phospho-STAT3 (CST, 9145, 1:100 dilution), and GR1 (R&D Systems, MN, USA, MAB1037-100, 1:100 dilution) overnight at 4 °C.

Techniques: Irradiation, Flow Cytometry, Immunofluorescence, Staining, Control