Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A The expression of neutrophils and CD8 + T cells in low- and high-glycolysis clusters, divided by ssGSEA score of glycolysis-related genes, in TCGA and CPTAC PAAD database. Data are presented as box plots showing the median (center line), the first and third quartiles (box bounds), and the whiskers extend to 1.5 times the interquartile range from the box. B The correlation of glycolysis level and neutrophils or CD8 + T cells expression in TCGA ( n = 179 patients) and CPTAC ( n = 140 patients) PAAD database. C Kaplan–Meier plots representing survival probabilities in TCGA-PAAD patients according to the relative level of glycolysis-related or neutrophil-related gene expression. D The diagram illustrating the targets of 2DG and shRNA in the glycolysis process. E CyTOF analysis of tumor-infiltrating immunocytes in subcutaneous tumor models with or without 2DG treatment: tSNE plots showing 12 meta-clusters based on the expression of 41 markers for the immunocytes. F Bar chart of the frequencies of the immune cell subsets in two experimental groups. G A schematic diagram showing the orthotopic PDAC model ( n = 6 mice per group in one experiment) with or without 2DG treatment. H , I Image and weights of the orthotopic tumors in the experimental groups from ( G ) at the end of the experiments. ( n = 6 mice per group). ( J – M ) Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). N A schematic diagram showing the orthotopic tumor model in C57BL/6 J mice ( n = 6 mice per group in one experiment) treated with or without anti-Ly6G/anti-CD8 antibody. O , P Image and weights of the orthotopic tumors in the experimental groups from ( N ) at the end of the experiments ( n = 6 mice per group). Q – S Tumor-infiltrating neutrophils, CD8 + T cells, and GZMB + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). T , U Representative luminescence images of the mouse model in ( N ) and statistical analysis of the total flux ( n = 6 mice per group). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( A, I–M, P–S, and U ), two-tailed Pearson’s correlation analysis ( B ), and log-rank test ( C ). Source data are provided as a Source Data file.

Article Snippet: Then, the mice were subjected to different treatments as shown in the figures and described as following: (1) Vehicle/2DG treatment was administered (800 mg/kg, i.p., once daily) from day 7 for 9 consecutive days; (2) For neutrophil depletion, these mice received anti-IgG2a (0.25 mg/mouse, i.p.) or anti-Ly6G antibody (0.25 mg/mouse, i.p., BioXCell, #BE0075-1) treatment once a day from day 3 post-implantation.

Techniques: Expressing, Gene Expression, shRNA, Isolation, Flow Cytometry, Two Tailed Test

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

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: