Journal: Journal of the National Cancer Center

Article Title: Overexpression of ornithine decarboxylase 1 mediates the immune-deserted microenvironment and poor prognosis in diffuse large B-cell lymphoma

doi: 10.1016/j.jncc.2024.10.001

Figure Lengend Snippet: In vitro experiments demonstrate the impact of ODC1 on macrophage polarization. (A) Flow cytometry analysis of CD206 + M2 macrophages and CD86 + M1 macrophages after co-culture of M0 THP-1 cells with DLBCL cell lines overexpressing or knocking down ODC1. (B) ELISA measurement of cytokine levels in the supernatants after co-culture with different DLBCL cell lines. (C) RT-PCR analysis of M1/M2 marker gene expression in macrophages after co-culture with different DLBCL cell lines. (D) Plasmid-mediated overexpression and knockdown of ODC1 in THP-1 cell lines, with western blotting (upper panel) and RT-PCR (lower panel) used to detect ODC1 protein expression and mRNA levels. (E) Flow cytometry analysis of CD206 + M2 macrophages and CD86 + M1 macrophages in THP-1 cells with ODC1 expression discrepancy. (F) ELISA measurement of cytokine levels in the supernatants from THP-1 cells with ODC1 expression discrepancy. (G) RT-PCR analysis of M1/M2 marker gene expression in macrophages from different groups. The unpaired t -test was used to assess statistical differences between two groups. * P < 0.05; ⁎⁎ P < 0.01; ⁎⁎⁎ P < 0.001; ⁎⁎⁎⁎ P < 0.0001.

Article Snippet: Subsequently, AF488 conjugated anti-human CD68 antibody (clone # Y1/82A, Elabscience), APC conjugated anti-mouse CD206 antibody (clone # C068C2, Elabscience), or PE conjugated anti-human CD86 antibody (clone # BU63, Elabscience) were added, and the cells were incubated in the dark at room temperature for 30 min. After centrifugation, the cells were resuspended in 200 μL of PBS with 1 % BSA and analyzed using flow cytometry.

Techniques: In Vitro, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Marker, Gene Expression, Plasmid Preparation, Over Expression, Knockdown, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: SIRPB1 regulates inflammatory factor expression in the glioma microenvironment via SYK: functional and bioinformatics insights

doi: 10.1186/s12967-024-05149-z

Figure Lengend Snippet: A , B SIRPB1 expression in astrocytoma and glioblastoma cell clusters. C Correlation of SIRPB1 with immune cell infiltration. D Association between macrophage infiltration and SIRPB1; Wilcoxon and Spearman tests. E SIRPB1 and macrophage correlation in TCGA-GBM via TIMER2.0. F Kaplan–Meier curves for OS of TCGA-GBMLGG with ICR-high. G Co-localization of SIRPB1 with TMEM119, CD86, and CD163 in glioma. H SIRPB1 levels in glioma and monocyte lines from CCLE. I SIRPB1 expression in human and mouse glioma, microglia, and monocyte lines

Article Snippet: The induced THP-1 macrophages were collected, Fc receptors were blocked by human Fc Receptor Blocking Solution (Maokangbio), stained with FITC Anti-Mouse/Human CD11b Antibody (Elabscience, clone:M1/70), 7-AAD (Elabscience), APC Anti-Human CD206 Antibody (Elabscience, clone:15–2) and PE Anti-Human CD86 Antibody (Elabscience, clone:BU63), and detected and analyzed by Beckman cytoflex flow cytometry.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: SIRPB1 regulates inflammatory factor expression in the glioma microenvironment via SYK: functional and bioinformatics insights

doi: 10.1186/s12967-024-05149-z

Figure Lengend Snippet: SIRPB1 Knockout and Macrophage Polarization. A T7E1 assay results. WT wild-type, NC negative control, PC positive control. B SIRPB1 and FLAG-cas9 expression in THP-1 lines. C Sanger sequencing of SIRPB1 WT and SIRPB1 KO . D Protein expression post-M1/M2 treatments. E mRNA levels of M1/M2 markers (* P < 0.05, ** P < 0.01, *** P < 0.001, Dunnett’s test). F Flow cytometry of CD11b, CD86, CD206 in THP-1 lines

Article Snippet: The induced THP-1 macrophages were collected, Fc receptors were blocked by human Fc Receptor Blocking Solution (Maokangbio), stained with FITC Anti-Mouse/Human CD11b Antibody (Elabscience, clone:M1/70), 7-AAD (Elabscience), APC Anti-Human CD206 Antibody (Elabscience, clone:15–2) and PE Anti-Human CD86 Antibody (Elabscience, clone:BU63), and detected and analyzed by Beckman cytoflex flow cytometry.

Techniques: Knock-Out, Negative Control, Positive Control, Expressing, Sequencing, Flow Cytometry

Journal: Journal of biomedical materials research. Part A

Article Title: DENDRITIC CELL RESPONSES TO SELF-ASSEMBLED MONOLAYERS OF DEFINED CHEMISTRIES

doi: 10.1002/jbm.a.32487

Figure Lengend Snippet: Dendritic cells treated with different SAMs exhibited differential expression levels of DC maturation markers. Dendritic cells matured with LPS exhibited higher expression of CD80, CD83 or HLA-DQ compared to iDCs. Dendritic cells treated with OH, COOH or NH2 SAMs increased expression of CD80, CD83 or CD86 as compared to iDCs, but to a lower extent than DCs treated with LPS as observed with CD80 expression. In contrast, DCs treated with CH3 SAMs had higher expression of HLA-DQ as compared to iDCs. This experiment was performed using six independent donors in triplicate. Data represented as treatment control ratios for each treatment revealed statistical significance of the findings; mean ± SEM, of pooled average ratios for each donor from n=6 donors; ‘+’ indicates increase over iDC, differences between SAMs in words, p≤0.05.

Article Snippet: Loosely adherent cell fractions containing DCs for iDCs, mDCs or DCs treated with different SAMs were collected and resuspended in Hank’s HEPES buffer (120 mM NaCl, 10 mM KCl, 10 mM MgCl 2 , 10 mM glucose, 30 mM HEPES) (all from Sigma) containing 1% Human Serum Albumin (HSA) (EMD Biosciences, San Diego, CA) and 1.5 mM CaCl 2 (Sigma) and stained with saturating concentrations of fluorescently-labeled mouse anti-human monoclonal antibody against CD14 (clone UCHM1; IgG2aκ), CD40 (clone B-B20; IgG1κ), CD80 (clone BB1; IgMκ), CD86 (clone BU63; IgG1κ), CD19 (clone SJ25-C1; IgG 1 ), CD14 (clone UCHM-1; IgG2a) (all from Southern Biotech, Birmingham, AL), CD83 (clone HB15a; IgG2b) (IO Test Immunotech Beckman Coulter, Marseille, France) HLA-DQ (clone TU169; IgG2aκ), HLA-DR (clone TU36; IgG2aκ), CD3 (clone SK7; IgG1κ), CD24 (clone ML5; IgG2aκ) (all from Becton Dickinson Pharmingen, San Diego, CA), CD1c (clone AD5–8E7; IgG2a) (Miltenyi Biotec, Auburn, CA), or cytotoxic T lymphocyte associated antigen receptor-4 (CTLA-4-Ig) (clone 48815; IgG 2b ) (R&D Systems, Minneapolis) while maintained on ice and in the dark.

Techniques: Quantitative Proteomics, Expressing, Control