Journal: Frontiers in Pharmacology

Article Title: Jianpi Yangzheng Xiaozheng decoction alleviates gastric cancer progression via suppressing exosomal PD-L1

doi: 10.3389/fphar.2023.1159829

Figure Lengend Snippet: JPYZXZ reduced the number of MDSCs in tumor bearing mice. (A,B) Percentage of MDSCs (CD45 + CD11b+Gr1+) in blood determined using flow cytometry. (C,D) Percentage of MDSCs (CD45 + CD11b+Gr1+) in tumors determined using flow cytometry. (E) Immunofluorescence staining for DAPI (nucleus), CD11b, Gr-1, and PD-L1 in tumor tissue sections from the xenograft tumor model. Data are represented as the mean ± SEM, n = 5 mice or n = 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The single-cell suspensions (1 × 10 7 cells/mL) were stained with fluorochrome-coupled antibodies AF488-CD45 (Elabscience, E-AB-F1136L), PE/Dazzle™594-CD11b (BioLegend, 101256), and APC-Gr-1 (BioLegend, 108412) for 30 min at 4°C.

Techniques: Flow Cytometry, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: APC-Vio770 Recombinant human anti B220(clone REA755) , Miltenyi Biotec , Cat# 130–110-849; RRID:AB_2658286.

Techniques: Negative Control, Recombinant, Purification, Staining, Gene Expression, Lysis, Immunoprecipitation, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Cell Isolation, Software, Microscopy

Journal: Cell reports

Article Title: Activation of GPR44 decreases severity of myeloid leukemia via specific targeting of leukemia initiating stem cells

doi: 10.1016/j.celrep.2023.112794

Figure Lengend Snippet:

Article Snippet: Mouse anti-mouse CD45.1 (FITC) , Miltenyi Biotec , Cat# 130–124-211; RRID: AB_2857674.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Binding Assay, Staining, Modification, Saline, Concentration Assay, Over Expression, Protein Extraction, Membrane, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Protease Inhibitor, CCK-8 Assay, Reverse Transcription, Selection, Plasmid Preparation, Knock-Out, Software, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: Nature Communications

Article Title: An in-situ peptide-antibody self-assembly to block CD47 and CD24 signaling enhances macrophage-mediated phagocytosis and anti-tumor immune responses

doi: 10.1038/s41467-024-49825-6

Figure Lengend Snippet: a Representative H&E and IF staining of cytokeratin 19 (CK19), F4/80 + macrophages and CD8 + T cells for the corresponding 4T1 tumor tissues after different treatments. Scale bars are marked in the figures. b Representative flow cytometry plots depicting CD45 + CD11b + F4/80 + macrophages phagocytosis, CD11b + F4/80 + CD206 hi macrophages, CD11b + F4/80 + CD86 hi macrophages, CD45 + CD3 + CD4 + /CD8 + T cells, and CD45 + CD3 + CD4 + Foxp3 + T cells in 4T1 tumors after different treatments. c – g Quantification analysis of CD45 + CD11b + F4/80 + macrophages phagocytosis, CD11b + F4/80 + CD206 hi macrophages, CD11b + F4/80 + CD86 hi macrophages, CD45 + CD3 + CD4 + /CD8 + T cells, and CD45 + CD3 + CD4 + Foxp3 + T cells in 4T1 tumors after different treatments. The error bars represent the mean ± SD ( n = 4 independent experiments; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; the p value was analyzed by a two-tailed unpaired Student’s t test). h Identification of differentially expressed genes in 4T1 tumors after treating with IgG Control or PAC-SABIs ( n = 5 mice). i Number of differential genes enriched in GO term of 4T1 tumors treated with PAC-SABIs versus IgG Control ( n = 5 mice). j GO enrichment analysis of the differential pathways in 4T1 tumors treated with PAC-SABIs versus IgG Control ( n = 5 mice). k Volcano plot for the transcriptome sequencing of 4T1 tumors treated with PAC-SABIs versus IgG Control. Differentially expressed genes were calculated using a two-sided limma moderated t -test with Benjamini–Hochberg correction for multiple testing ( n = 5 mice). Source data are provided as a Source Data file.

Article Snippet: Flow cytometry: PerCP/Cy5.5 anti-mouse CD45 antibody (1:200, Elabscience, #E-AB-F1136J, clone 30-F11), PE/Cyanine5 anti-mouse CD11b antibody (1:200, Biolegend, #101209, clone M1/70), APC anti-mouse F4/80 antibody (1:200, Biolegend, #123116, clone BM8), FITC anti-mouse CD206 antibody (1:200, Biolegend, #141703, clone C068C2), FITC anti-mouse CD86 antibody (1:200, Biolegend, #159219, clone A17199A), FITC anti-mouse CD3 antibody (1:200, BDbioscience, #555274, clone 17A2), APC anti-mouse CD4 antibody (1:200, BDbioscience, #553051, clone RM4-5), PE anti-mouse CD8a antibody (1:200, BDbioscience, #553033, clone 53-6.7), PE anti-mouse Foxp3 antibody (1:200, BDbioscience, #563101, clone R16-715).

Techniques: Staining, Flow Cytometry, Two Tailed Test, Control, Sequencing

Journal: Nature Communications

Article Title: An in-situ peptide-antibody self-assembly to block CD47 and CD24 signaling enhances macrophage-mediated phagocytosis and anti-tumor immune responses

doi: 10.1038/s41467-024-49825-6

Figure Lengend Snippet: a Scheme of the combination therapeutic strategy. Figure was created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 international license. b Spider plots of individual tumor growth curves for 4T1-tumor-bearing BALB/c mice in each treatment group. c Average tumor growth curve of 4T1-tumor-bearing BALB/c mice in each treatment group. The error bars represent the mean ± SD ( n = 7 mice; **** p < 0.0001; the p value was analyzed by a two-tailed unpaired Student’s t test). d Kaplan–Meier survival curves of 4T1 tumor-bearing BALB/c mice treated with the indicated formulation. ( n = 7 mice; ** p < 0.01, *** p < 0.001; the p value was analyzed by log-rank test). e Representative H&E staining of lungs harvested from the indicated treatment group. Scale bar: 100 μm. Three independent experiments were performed. f Quantitative analysis of phagocytosis in CD45 + CD11b + F4/80 + macrophages flow cytometry results. The error bars represent the mean ± SD ( n = 4 independent experiments; *** p < 0.001, **** p < 0.0001; the p value was analyzed by a two-tailed unpaired Student’s t test). g Representative IF images of tumor-infiltrated CD8 + T cells. Scale bar: 100 μm. Three independent experiments were performed. Source data are provided as a Source Data file.

Article Snippet: Flow cytometry: PerCP/Cy5.5 anti-mouse CD45 antibody (1:200, Elabscience, #E-AB-F1136J, clone 30-F11), PE/Cyanine5 anti-mouse CD11b antibody (1:200, Biolegend, #101209, clone M1/70), APC anti-mouse F4/80 antibody (1:200, Biolegend, #123116, clone BM8), FITC anti-mouse CD206 antibody (1:200, Biolegend, #141703, clone C068C2), FITC anti-mouse CD86 antibody (1:200, Biolegend, #159219, clone A17199A), FITC anti-mouse CD3 antibody (1:200, BDbioscience, #555274, clone 17A2), APC anti-mouse CD4 antibody (1:200, BDbioscience, #553051, clone RM4-5), PE anti-mouse CD8a antibody (1:200, BDbioscience, #553033, clone 53-6.7), PE anti-mouse Foxp3 antibody (1:200, BDbioscience, #563101, clone R16-715).

Techniques: Two Tailed Test, Formulation, Staining, Flow Cytometry