Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Interleukin (IL)-18 amplifies macrophage (Mφ) M2 polarization and angiogenic capacity. (A) Representative FACS density plots for the expression of CD86 and CD163 in each Mφ subset. Upper, Mφ (–); middle; Mφ [tumor necrosis factor (TNF)-α]; lower, Mφ (IL-10). The numbers in each quartile of the plots are percentages of each cell population. (B) Relative mean fluorescence intensities (MFIs) of CD54, CD86, CD163, and CD206 in each Mφ subset were measured by FACS analysis, n = 3 (*** p < 0.001, * p < 0.05 vs. untreated, ## p < 0.01 vs. IL-10 alone). (C) Relative MFI of IL-18Rβ in each Mφ subset was determined by FACS analysis, n = 4 (*** p < 0.001 vs. untreated, ### p < 0.001 vs. IL-10 alone). (D,E) The total areas and lengths of tube-like structures measured by the Matrigel tube formation assay where b.End5 was cocultured with each Mφ subset, n = 6 (*** p < 0.001, ** p < 0.01, * p < 0.05 vs. untreated, ## p < 0.01, # p < 0.05 vs. IL-10 alone). (F) Representative pictures of tube-like structures visualized by calcein acetoxymethylester staining. Scale bar represents 100 µm. All data are presented as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 μL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Expressing, Fluorescence, Tube Formation Assay, Staining

Journal: Frontiers in Immunology

Article Title: Paeonol Interferes With Quorum-Sensing in Pseudomonas aeruginosa and Modulates Inflammatory Responses In Vitro and In Vivo

doi: 10.3389/fimmu.2022.896874

Figure Lengend Snippet: Paeonol changed the Polarization of Macrophages infected with PAO1 (MOI = 25:1). Control (A) , PAO1 (B) The data in “PAO1+Pae 32 μg/mL” (D) , “PAO1+Pae 64 μg/mL” (E) , and “PAO1+Pae 128 μg/mL” (F) indicated that Paeonol reversed the upregulated the biomarker of CD86 of RAW264.7 cells infected with PAO1 (C) . The cell polarization was distinguished by Flow cytometry. All data were expressed as means ± SD (n=3). * P < 0.05 or ** P < 0.01 vs PAO1 group.

Article Snippet: The cells were stained with F4/80 (Elabscience, E-AB-F0995C), CD86 (Elabscience, E-AB-F0994E), and CD206 (Elabscience, E-AB-F1135D) fluorescently labeled antibody, then detected by a flow cytometer (Beckman coulter, CytoFLEX) and analyzed by the Flowjo software.

Techniques: Infection, Control, Biomarker Discovery, Flow Cytometry

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: Fig. 2 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: Fig. 4 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 SIRPB1WT and SIRPB1KO. 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: Materials Today Bio

Article Title: 4D-printed Fe3O4-PLLA scaffolds modulate osteoimmune microenvironment for oral bone repair

doi: 10.1016/j.mtbio.2025.102691

Figure Lengend Snippet: In vitro immunomodulatory properties of PLLA, Fe 3 O 4 -PLLA, CSMA/Fe 3 O 4 -PLLA, NG-CSMA/Fe 3 O 4 -PLLA scaffolds. a-b) 3 days of co-culture of RAW 264.7 with PLLA, Fe 3 O 4 -PLLA, CSMA/Fe 3 O 4 -PLLA, and NG-CSMA/Fe 3 O 4 -PLLA scaffolds, flow cytometry was performed to analyze M1, M2 markers CD86 and CD206. c) mRNA levels of TNF-α, IL-1β, Arg-1, and CD206 by qRT-PCR; d) Immunofluorescence images of M1 (iNOS) and M2 (Arg-1) macrophage markers; and e) quantitative analysis of fluorescence intensity. n = 3. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).

Article Snippet: After 3 days, cells were harvested, blocked, and stained with PE-anti-CD206 and CL647- anti -CD86 antibodies (Proteintech, China).

Techniques: In Vitro, Co-Culture Assay, Flow Cytometry, Quantitative RT-PCR, Immunofluorescence, Fluorescence