Journal: American Journal of Translational Research

Article Title: Adoptive transfer of polarized M2c macrophages ameliorates acute rejection in rat liver transplantation

doi:

Figure Lengend Snippet: Primers used for qRT-PCR

Article Snippet: Primary anti-rat CD163 antibody (NBP2-39099, Novus Biologicals Europe) was used at a dilution of 1:100; primary anti-rat MHC-II antibody (ab23990, Abcam) was used at a dilution of 1:300, whereas the PBS was used as a negative control.

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: Adoptive transfer of polarized M2c macrophages ameliorates acute rejection in rat liver transplantation

doi:

Figure Lengend Snippet: The infiltration of the CD163-positive cells increased in the tolerant liver grafts. A. Representative images of HE staining of liver allografts in the tolerance and the AR group 7 days following transplantation with original magnifications of ×100. Characteristics like portal inflammatory cell infiltration, bile duct damage and endothelial inflammation significantly reduced in the tolerance group. Scale bar in right lower corner represents 100 µm. B. Liver functions were assessed on day 7 after transplantation. Both ALT and AST were significantly lowered in tolerance group. C, D. ELISA was used to detect serum IL-10 and TGF-β1 levels of the recipients in both groups. Both anti-inflammatory cytokines were significantly increased in the tolerant recipients. E. Illustrating IHC microscopic finding for identification of CD163 positive cells (brown color) with original magnifications of ×400. Scale bar in right lower corner represents 25 µm. F. Analytical results of the numbers of CD163 positive cells. The numbers of CD163 positive cells in the AR group were less than that in the tolerance group. All statistical analyses were performed by an unpaired t-test. Data are presented as the mean ± SD. (n = 5, *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Primary anti-rat CD163 antibody (NBP2-39099, Novus Biologicals Europe) was used at a dilution of 1:100; primary anti-rat MHC-II antibody (ab23990, Abcam) was used at a dilution of 1:300, whereas the PBS was used as a negative control.

Techniques: Staining, Transplantation Assay, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: Adoptive transfer of polarized M2c macrophages ameliorates acute rejection in rat liver transplantation

doi:

Figure Lengend Snippet: M2c BMDMs were successfully induced in vitro. A. The bone marrow derived cells were examined by immunofluorescence staining with anti-CD68 antibody after being stimulated by MCSF for 7 days. Nearly all cells expressed CD68, the specific rat macrophage marker (Magnification, 200). Scale bars in right lower corner represents 50 µm. These cells were then stimulated by PBS or dexamethasone for 24 h for M0 or M2c polarization. B. Representative images of the M0 and the M2c with original magnifications of ×400. Scale bar in right lower corner represents 25 µm. C. M2c polarization markers expression determined by qRT-PCR. The expression levels of CD163, IL-10, TGF-β1 in the M2c macrophages were significantly higher than those of the M0 macrophages. The statistical analyses were performed by an unpaired t-test. Data are presented as the mean ± SD. (n = 3, **P < 0.01).

Article Snippet: Primary anti-rat CD163 antibody (NBP2-39099, Novus Biologicals Europe) was used at a dilution of 1:100; primary anti-rat MHC-II antibody (ab23990, Abcam) was used at a dilution of 1:300, whereas the PBS was used as a negative control.

Techniques: In Vitro, Derivative Assay, Immunofluorescence, Staining, Marker, Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Regenerative Effects of Hypoxia Primed Flowable Placental Formulation in Muscle and Dermal Injury

doi: 10.3390/ijms22137151

Figure Lengend Snippet: Angiogenic and anti-inflammatory response in FPF-treated wounds. ( A ) Neovascularization in control (left panel) and FPF treated (right panel) tissue samples collected after wound closure was determined by αSMA (top panel, stained green) and CD31 (middle panel, stained red). Merged images (bottom panel) show the colocalization of αSMA and CD31. Nuclei were counterstained with DAPI (blue). White arrows indicate αSMA- and CD31-positive blood vessels. Graphs represent the vessel density and size distribution of αSMA and CD31positive blood vessels. Scale bar = 50 μm. N = 6 for saline; N = 9 for FPF group. Inflammatory cell staining was determined by ( B ) MPO (red) staining for neutrophils. Scale bar = 50 μm and ( C ) CD163 (red) staining for M2 macrophages. Scale bar = 50 μm. Nuclei were counterstained with DAPI (blue). N = 6/group. * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: The following primary antibodies and concentrations were used: for α smooth muscle actin (αSMA), rabbit polyclonal anti-αSMA primary antibody for 45 min (0.33 μg/mL) (#ab5694, Abcam, Waltham, MA, USA); for CD31, mouse monoclonal (TLD-3A12) anti-CD31 primary antibody for 45 min (10 μg/mL) (#MA1-80069, Invitrogen, Waltham, MA, USA); for CD68, mouse monoclonal (ED-1) anti-CD68 primary antibody for 30 min (5 μg/mL) (#MCA341, Biorad, Hercules, CA, USA); for CD163, mouse monoclonal (ED-2) anti-CD163 primary antibody for 30 min (6.67 μg/mL) (#MCA342, Biorad, Hercules, CA, USA); for collagen IV, rabbit polyclonal anti-collagen IV primary antibody for 60 min (10 μg/mL) (Invitrogen #PA1-28534); for MPO, rabbit polyclonal anti-MPO primary antibody for 30 min (1.33 μg/mL) (#ab9535, Abcam, Waltham, MA, USA).

Techniques: Control, Staining, Saline

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