Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: All primary and secondary antibodies used in this study.

Article Snippet: Mouse MMR/CD206 APC-conjugated Antibody , Goat , , 1:20 , R & D Systems , FAB2535A.

Techniques:

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet: Combination of CITE-seq, scRNA-seq, snRNA-seq, and spatial analyses enables identification of all hepatic cell types including bona fide cell doublets, related to Figure 1 (A and B) Top DEGs (A) and DEPs (B) for cell types from Figure 1 B. (C) Distinct profiles of cells or nuclei within the UMAP depending on isolation protocols; 71,162 cells from ex vivo digestions, 96,066 cells from in vivo digestions, and 18,666 nuclei. Numbers on plots represent numbers of cells/nuclei per population. (D) Correlation plots showing genes captured within the KC, B cell and neutrophil populations with and without addition of CITE-seq antibodies. (E) Expression of VSIG4, CD206, and ESAM (protein, top) and Vsig4 , Mrc1 , and Esam (mRNA, bottom). (F) UMAP showing clusters of cells when only minimal QC for gene number and % mitochondrial genes is performed; 17,669 cells pooled from 3 samples. Expression of Cd5l, Cd19 , and Kdr by the clusters facilitating identification of cell types per annotation. (G) CITE-seq data from (F) in Flow-Jo showing expression of CD206 and ESAM in total KCs (left) and total B cells (middle). Numbers represent % of entire KC or B cell population. Identified populations were then mapped back onto the original UMAP (right). (H) Expression of CD31, CD26, and CD38 by indicated populations. (I) Heatmaps showing expression of top DEGs between KC1s and LSECs (left), KC2s and KC1s + LSECs (middle) and B cell2s and B cell1s + LSECs (right). (J) 3D reconstruction of murine liver following perfusion with antigen fix to inflate endothelial cells and staining with antibodies against CD31, CD206, and F4/80. (K) UMAP showing clusters generated from Visium analysis of liver tissue (4 samples) and liver capsule (1 sample). (L) Top unbiased genes defining zonation trajectory from portal to central vein in Visium. (M) Expression of Glul and Epcam by confocal microscopy (left), annotation of portal, periportal, mid, and central regions on same tissue section (middle) and overlay of both datasets (right). (N) Identification of cholangiocyte (left) and cDC (right) signatures on zonated Visium spots. (P) Molecular Cartography showing expression of indicated zonated hepatocyte mRNAs in liver tissue. Data are representative of 2 mice. (O) Expression of Itgae (encoding CD103) in the UMAP of the total liver (left) and flow cytometric analysis of total cDC1s for CD103 and MHCII expression in the healthy murine liver (right).

Article Snippet: Anti-Human CD206 (REAL518) APC , Miltenyi Biotec , 130-122-168; RRID: AB_2857557.

Techniques: Isolation, Ex Vivo, In Vivo, Expressing, Staining, Generated, Confocal Microscopy

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Human CD206 (REAL518) APC , Miltenyi Biotec , 130-122-168; RRID: AB_2857557.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

Journal: Chemical Engineering Journal

Article Title: Cryogenically 3D printed biomimetic scaffolds containing decellularized small intestinal submucosa and Sr2+/Fe3+ co-substituted hydroxyapatite for bone tissue engineering

doi: 10.1016/j.cej.2021.133459

Figure Lengend Snippet: Fig. 5. In vitro RAW cells polarization modulation after cultured on scaffolds for 3 days. (A) Calcein-AM staining of RAW; (B) RAW cells polarization examined by double marked immunofluorescence of NOS2/Arg-1; (C) RAW polarization quantified by double marked flow cytometry of CD86/CD206; (D) Quantitative CD206 and CD86 expression ratios; (E) RT-qPCR analysis of relative IL6, IL1-β, TNF-α, Arg-1 and IL10 gene expressions; (F) RT-qPCR analysis of relative VEGF, PDGF-BB, TGF-β1 and BMP2 gene expressions. Data represent the mean ± SD. *p < 0.05 versus SIS; #p < 0.05 versus SIS/FeHA.

Article Snippet: Briefly, RAW cells were harvested and incubated with CD86 antibody (553692, BD Pharmingen, USA) under 4 °C for 30 min. Then, cells were washed, resuspended, and incubated with CD206 antibody (FAB25351A, R&D Systems, USA) again to be analyzed using the flow cytometer (Beckman Coulter, CytoFLEX).

Techniques: In Vitro, Cell Culture, Staining, Immunofluorescence, Flow Cytometry, Expressing, Quantitative RT-PCR

Journal: Chemical Engineering Journal

Article Title: Cryogenically 3D printed biomimetic scaffolds containing decellularized small intestinal submucosa and Sr2+/Fe3+ co-substituted hydroxyapatite for bone tissue engineering

doi: 10.1016/j.cej.2021.133459

Figure Lengend Snippet: Fig. 7. In vivo immunomodulation and vascularization of scaffolds following 7 days subcutaneous implantation. (A) The histological graphs of implanted scaffolds : from the general view of horizontal HE sections to internal sites stained by HE, NOS2, CD206, and CD31 IHC in similar positions; (B). Quantitation of CD206 and NOS2 positive stained areas; (C) Quantitative CD206/NOS2 ratios; (D) Quantitation of new vessels density. Red arrows indicate neo-vessels stained by CD31. Data represent the mean ± SD. *p < 0.05 versus SIS; &p < 0.05 versus SIS/FeHA; #p < 0.05 versus SIS/SrHA.

Article Snippet: Briefly, RAW cells were harvested and incubated with CD86 antibody (553692, BD Pharmingen, USA) under 4 °C for 30 min. Then, cells were washed, resuspended, and incubated with CD206 antibody (FAB25351A, R&D Systems, USA) again to be analyzed using the flow cytometer (Beckman Coulter, CytoFLEX).

Techniques: In Vivo, Staining, Quantitation Assay

Journal: Biomaterials Research

Article Title: Custom-Made Ce–Mn Bimetallic Nanozyme for the Treatment of Intervertebral Disc Degeneration by Inhibiting Oxidative Stress and Modulating Macrophage M1/M2 Polarization

doi: 10.34133/bmr.0118

Figure Lengend Snippet: CeMn-PEG could promote the polarization of macrophages to M2 type. (A to C) Representative images and relative fluorescence intensity of CD86 and CD206 in RAW264.7 cells in the coculture system. (D and E) Representative images of cytometry analysis (D) and mean fluorescence intensity (MFI) quantification (E) of CD86 and CD206 in RAW264.7 cells in the coculture system. (F and G) Representative images and quantification data of Western blot results of IL-1β, IL-4, IL-6, and IL-10 in RAW264.7 cells in the coculture system. Scale bar, 25 μm. Data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n = 3.

Article Snippet: After NP and RAW264.7 cells were treated as previously described, the digested single-cell suspension was incubated with flow cytometry antibodies, CD86-FITC (fluorescein isothiocyanate), and CD206-APC (allophycocyanin) at room temperature in the dark for 1 h. The V-APC/7-AAD Apoptosis Detection Kit (KeyGEN) was used according to the manufacturer’s instruction.

Techniques: Fluorescence, Cytometry, Western Blot