Journal: Acta biomaterialia

Article Title: A subtype specific probe for targeted magnetic resonance imaging of M2 tumor-associated macrophages in brain tumors.

doi: 10.1016/j.actbio.2025.01.003

Figure Lengend Snippet: Fig. 6. Immunohistochemical and FACS validations of M2pep-uIONP targeting M2 TAMs. Confocal microscopic images of tumor tissues collected from mice at 24 h after i.v. injection of probes show (A) Cy7-M2pep-uIONP were highly co-localizing with CD68- and CD163-double positive M2 TAM, revealing the targeting specificity. In comparison, the non-targeted control probe Cy7-scM2pep-uIONP and Cy7-Ferumoxytol not only exhibited much less tumoral accumulation, but also were not co-localized with M2 TAM. Scale bars: 37 µm. (B) Flow cytometric zebra plots and (C) the corresponding quantification of CD68- and CD163-double positive M2 TAM targeted by FITC-M2pep-uIONP, FITC-scM2pep-uIONP, and FITC-Ferumoxytol. The gating strategy involves sorting CD163+ M2 TAM population in live cells dissociated from mouse cerebrums by gating on CD68+ pan-macrophage populations, followed by further gating to assess the FITC-labeled nanoparticles.

Article Snippet: Rabbit polyclonal anti-CD163 antibody was purchased from Boster Biological Technology (Pleasanton, CA, USA).

Techniques: Immunohistochemical staining, Injection, Comparison, Control, Labeling

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: IL-13–regulated Macrophage Polarization during Granuloma Formation in an In Vitro Human Sarcoidosis Model

doi: 10.1165/rcmb.2018-0053oc

Figure Lengend Snippet: Figure 5: Immunofluorescence imaging of granuloma-like structures demonstrates persistent macrophage CD163 expression on sarcoidosis PBMCs following PPD stimulation. Representative photomicrographs of immunostained PBMCs in granuloma-like structures 7 days after bead treatment using

Article Snippet: Copyright © 2018 by the American Thoracic Society Alternatively, following washing and blocking, some coverslips were incubated with 0.5 μg/ml CD163 [unlabeled, rabbit, anti-human CD163 mAb (K20-T, IgG) from Novus Biologicals, LLC (Littleton, CO)] or the matching isotype control antibody [Abcam, (Cambridge, MA)] at room temperature for 1 hour.

Techniques: Immunofluorescence, Imaging, Expressing

Journal: Cell Death & Disease

Article Title: M2 macrophage-mediated interleukin-4 signalling induces myofibroblast phenotype during the progression of benign prostatic hyperplasia

doi: 10.1038/s41419-018-0744-1

Figure Lengend Snippet: a Representative CD68 immunohistochemical staining images in the early-progressed BPH, age-matched prostate and elderly BPH tissues. Scale bar, 200 μm. b Scatter plots showing macrophage clusters in the three analysed groups. The number of macrophage clusters in prostate tissues correlated positively with the stroma-to-epithelium ratio, and percent area densities of α-SMA, collagen I and fibres. c Serial histological sections showing the early-progressed BPH tissues stained for CD68, a macrophage-specific marker, and CD163, an M2 macrophage marker. The distribution of CD68 + and CD163 + cells was almost identical. Scale bar, 200 μm. d Representative immunofluorescence (IF) staining images, showing CD68 (green) and CD163 (red) expression in the infiltrating macrophages in early-progressed BPH tissues. Scale bar, 100 μm. e Quantitative RT-PCR results showing α-SMA, COL1A1 and COL3A1 expression in primary prostate fibroblast (PrPF)-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. Data are shown as relative gene expression compared to that in the respective untreated fibroblasts. f Western blots showing α-SMA and collagen I protein expression in PrPF-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. g Representative IF staining results, showing the co-expression of α-SMA (red) and collagen I (green) following a 48-h co-culture of PrPF-early, PrPF-control and PrPF-old cells with THP-1-derived M2 macrophages. The respective untreated fibroblasts were used as controls. Scale bar, 100 μm. h Solidified collagen-gel shrinkage after the seeding of PrPF-early, PrPF-control and PrPF-old cells, untreated or treated with the conditioned medium. The respective untreated fibroblast samples were used as controls, and all assays were performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies included mouse monoclonal α-SMA (Abcam, ab7817, 1:100 dilution), CD163 (OriGene, TA506380, 1:200 dilution) and EpCAM (Cell Signaling Technology, #2929, 1:800 dilution) antibodies, and the rabbit primary antibodies included collagen I (Abcam, ab138492, 1:800 dilution), CD68 (Proteintech, 25747-1-AP, 1:400 dilution), IL4R (ImmunoWay, YT2337, 1:400 dilution), TGFβ RI (ImmunoWay, YT4627, 1:400 dilution) and vimentin (Cell Signaling Technology, #5741, 1:100 dilution) antibodies.

Techniques: Immunohistochemical staining, Staining, Marker, Immunofluorescence, Expressing, Quantitative RT-PCR, Control, Cell Culture, Derivative Assay, Gene Expression, Western Blot, Co-Culture Assay