Journal: Glia

Article Title: Activated microglia do not increase 18 kDa translocator protein ( TSPO ) expression in the multiple sclerosis brain

doi: 10.1002/glia.24052

Figure Lengend Snippet: Antibodies for immunohistochemistry

Article Snippet: CD68 , Rabbit , Atlas Antibody , HPA048982 , 1:4000.

Techniques:

Journal: PLOS One

Article Title: Photobiomodulation therapy increases neural stem cell pool in aged 3xTg-AD mice

doi: 10.1371/journal.pone.0321668

Figure Lengend Snippet: A. Representative confocal images of Aβ plaques (6E10), microglia (Iba1) and CD68 showing diffuse microglial phagocytosis away from plaques in sham compared to localized phagocytosis around plaques in PBM-treated animals. B. Quantitative analysis of total and activated microglia show no overall differences in the neuroinflammatory environment surrounding Aβ plaques. (Unpaired t-test). C. Analysis of the distribution of CD68+ microglia shows a trend of higher concentration of phagocytic cells in contact with plaques, and decreasing phagocytosis away from plaques. (Two-way ANOVA).

Article Snippet: Up to six sections representing different regions of the hippocampus were immunostained with the following primary antibodies: Sox2 (Abcam), doublecortin (DCX, Santa Cruz), calretinin (CR, SWANT), β-amyloid (6E10, Biolegend), phospho-tau (pTau, AT8, Invitrogen), βIII-tubulin (Abcam), Iba1 (Synaptic Systems), and CD68 (Rockland).

Techniques: Concentration Assay

Journal: The American journal of pathology

Article Title: The Versican G1 Fragment and Serum-Derived Hyaluronan-Associated Proteins Interact and Form a Complex in Granulation Tissue of Pressure Ulcers.

doi: 10.1016/j.ajpath.2017.10.015

Figure Lengend Snippet: Figure 9 Transient versican G1 fragmenteserum-derived hyaluronan-associated proteinehyaluronan (VG1F-SHAP-HA) complex formation in a mouse model of moist wound healing. A: Representative images from wound tissues from the moist wound-healing model are shown. Edematous granulation tissues were observed at day 3 after wounding; however, the granulation tissues were nonedematous and thin at days 1 and 6. The thickness of the granulation tissues is as follows: day 1, 0.77 0.10 mm; day 3, 2.55 0.17 mm; and day 6, 0.82 0.04 mm. B: Tissue specimens were stained with hematoxylin and eosin. C: Isolated tissues were stained by using the indicated probes. The VG1F-SHAP-HA complex was observed on day 3 in specific lesions, but was absent at days 1 and 6. The color of letters indicating molecules represents the color of the stain. The merged images indicate the VG1F-SHAP-HA complex. D: Immunohis- tochemical analysis of wound tissue taken from day 3. The specimen was stained with the indicated probes, and the merged images show VG1F colocalization, as detected by probes for HA, DPEAAE epitope, and macrophages (CD68). The merged images show macrophage/complex colocalization. Data are expressed as means SD (A). n Z 4 independent experiments (A). Scale bars Z 50 mm (BeD). HC, heavy chain.

Article Snippet: A goat pAb against bikunin (sc-21597) was purchased from Santa Cruz Biotechnology Inc. A mouse monoclonal antibody against the macrophage marker CD68 (STJ-6572) was purchased from St. John’s Laboratory Ltd (London, UK).

Techniques: Derivative Assay, Staining, Isolation

Journal: The American journal of pathology

Article Title: The Versican G1 Fragment and Serum-Derived Hyaluronan-Associated Proteins Interact and Form a Complex in Granulation Tissue of Pressure Ulcers.

doi: 10.1016/j.ajpath.2017.10.015

Figure Lengend Snippet: Figure 11 The versican G1 fragment (VG1F)eserum-derived hyaluronan (HA)eassociated proteineHA complex in human pressure ulcers. A: In human pressure ulcers, edematous wounds (e1 and e2) typically become flat and stabilized (f3 and f4) after appropriate treatments. B: The VG1F-HC complex was detected from the protein extract of wound surfaces shown in A (e1 and e2). The samples are indicated at the top of the figure and correspond to those shown in A. The samples were extracted with 6 mol/L GdnHCl and treated with trypsin and V8 protease (Materials and Methods); separated using SDS-PAGE under nonreducing conditions; and blotted. Incubation with the indicated antibodies revealed the cross-linked heavy chain (HC) and VG1 species (dagger). The bands of cross-linked HC and VG1 species (dagger) disappeared during wound healing. C: Paired samples from edematous or flat wounds were analyzed using Western blotting, and the bands shown in B were quantified. The relative intensities of the bands [VG1-HCs (daggers in B)]/the total polyclonal antibody 6084epositive bands (all VG1) differ significantly (P Z 0.02) between edematous and flat wounds. D: Direct immunofluorescence of wound surface samples obtained from different sites of one individual wound. Samples obtained from the edematous wound (green dashed circle) and flat wound (blue dashed circle) were fixed and incubated with VG1 and HC1 antibodies, as well as biotin-conjugated HAebinding protein, as indicated. A representative result from triplicate experiments with different samples is shown. E: Immunohistochemical analysis of pressure ulcer tissue. The specimen was stained, as indicated. The merged images show VG1F colocalization, as detected by antibodies for DPEAAE epitope, HC1, and macrophage CD68. Data are expressed as means SD (C). n Z 10 (C, each paired sample). Scale bars: 5 mm (D); 100 mm (E).

Article Snippet: A goat pAb against bikunin (sc-21597) was purchased from Santa Cruz Biotechnology Inc. A mouse monoclonal antibody against the macrophage marker CD68 (STJ-6572) was purchased from St. John’s Laboratory Ltd (London, UK).

Techniques: Derivative Assay, SDS Page, Incubation, Western Blot, Immunohistochemical staining, Staining

Journal: BMC Nephrology

Article Title: RIPK3-deficient mice were not protected from nephrotoxic nephritis

doi: 10.1186/s12882-018-0850-4

Figure Lengend Snippet: Histology at day 8 after NTN induction. Representative histology from PAS stained sections from Experiment 2, of WT and RIPK3−/− kidney. a ) WT b ) RIPK3- /−. Immunoperoxidase staining for CD68 in WT and RIPK3−/− kidney showing glomerular macrophage infiltration following the induction of NTN. c ) WT d ) RIPK3−/−

Article Snippet: Sections were cut to a thickness of 5 μm, incubated with the anti- CD68 antibody and this was detected using the Polink-2 plus HRP rat detection kit (Newmarket Scientific, Newmarket, U.K.).

Techniques: Staining, Immunoperoxidase Staining

Journal: American Journal of Translational Research

Article Title: The distribution of macrophage subtypes and their relationship to bone morphogenetic protein 2 in calcified aortic valve stenosis

doi:

Figure Lengend Snippet: Representative immunostaining images showing the expression of macrophage subtype markers [CD68 (A), CD163 (B), CD206 (C), heme oxygenase-1 (D), and inducible nitric oxide synthase (iNOS) (E)] and bone morphogenetic protein 2 (BMP2) (F) by single immunohistochemistry in the valve center part and spongiosa layer of a single case in the calcification group. Cells of the iNOS+ M1 and CD163+/CD206+ M2 subtypes frequently increased, while cells of the CD68+ M1/M2 subtype and heme oxygenase-1+ Mox subtypes were relatively few. Note that scattered BMP2+ cells were also present. iNOS and BMP2 were present in some vascular endothelial cells.

Article Snippet: For IHC, formalin-fixed and paraffin-embedded tissue sections (4 μm in thickness) were blocked with 0.3% H 2 O 2 in methanol at 4°C for 30 min and then subjected to antigen retrieval in trypsin for 30 min at 37°C for anti-CD68 antibody, in citric acid (Antigen Retrieval Solution pH 6; Iatron Laboratories Inc., Tokyo, Japan) for anti-CD 163 antibody, anti-iNOS antibody, anti-HO-1 antibody, and anti-OPN antibody, and in EDTA (pH 9.0; Nichirei, Tokyo, Japan) in an autoclave (2 atmospheres, 121°C, 20 min) for anti-CD206 antibody.

Techniques: Immunostaining, Expressing, Immunohistochemistry

Journal: American Journal of Translational Research

Article Title: The distribution of macrophage subtypes and their relationship to bone morphogenetic protein 2 in calcified aortic valve stenosis

doi:

Figure Lengend Snippet: Frequency of cells immunopositive for macrophage subtype markers and bone morphogenetic factor-2 (BMP2) in the calcification and noncalcification groups

Article Snippet: For IHC, formalin-fixed and paraffin-embedded tissue sections (4 μm in thickness) were blocked with 0.3% H 2 O 2 in methanol at 4°C for 30 min and then subjected to antigen retrieval in trypsin for 30 min at 37°C for anti-CD68 antibody, in citric acid (Antigen Retrieval Solution pH 6; Iatron Laboratories Inc., Tokyo, Japan) for anti-CD 163 antibody, anti-iNOS antibody, anti-HO-1 antibody, and anti-OPN antibody, and in EDTA (pH 9.0; Nichirei, Tokyo, Japan) in an autoclave (2 atmospheres, 121°C, 20 min) for anti-CD206 antibody.

Techniques:

Journal: American Journal of Translational Research

Article Title: The distribution of macrophage subtypes and their relationship to bone morphogenetic protein 2 in calcified aortic valve stenosis

doi:

Figure Lengend Snippet: Evaluation of the expression of bone morphogenetic protein 2 (BMP2) in macrophage subtypes by double immunofluorescence staining. Because the anti-iNOS antibody and the anti-BMP2 antibody were both rabbit polyclonal antibodies, single immunofluorescence staining was performed for each of the two proteins using serial sections. Nuclei were counterstained with 4’,6-diamidino-2-phenylindole (blue). Co-expression of BMP2 (green) was confirmed in a subset of the cells that showed positive staining for CD68 (M1/M2 subtype), CD206 (M2 subtype), and heme oxygenase-1 (Mox subtype) (red). However, co-expression of iNOS (M1 subtype) and BMP2 was not observed.

Article Snippet: For IHC, formalin-fixed and paraffin-embedded tissue sections (4 μm in thickness) were blocked with 0.3% H 2 O 2 in methanol at 4°C for 30 min and then subjected to antigen retrieval in trypsin for 30 min at 37°C for anti-CD68 antibody, in citric acid (Antigen Retrieval Solution pH 6; Iatron Laboratories Inc., Tokyo, Japan) for anti-CD 163 antibody, anti-iNOS antibody, anti-HO-1 antibody, and anti-OPN antibody, and in EDTA (pH 9.0; Nichirei, Tokyo, Japan) in an autoclave (2 atmospheres, 121°C, 20 min) for anti-CD206 antibody.

Techniques: Expressing, Double Immunofluorescence Staining, Immunofluorescence, Staining

Journal: American Journal of Translational Research

Article Title: The distribution of macrophage subtypes and their relationship to bone morphogenetic protein 2 in calcified aortic valve stenosis

doi:

Figure Lengend Snippet: Scheme of a possible mechanism of aortic valve calcification. Noncalcified valves contain a few macrophages, predominantly those of the M1 subtype [inducible nitric oxide synthase (iNOS)+/CD68+] and, less frequently, cells of the M2 subtype (CD68+/CD163+/CD206+). In the pre-calcification phase, against a background of fibrosis, M1 and M2 subtypes gradually accumulate, with a mildly increasing ratio of cells of the M2 subtype to those of the M1 subtype. A few heme oxygenase (HO)-1+ Mox appear. In the calcified valve, calcification is accompanied by the presence of osteoblast-like cells (OLC). In this valve, all subtypes of macrophages are increasing, with a relatively high ratio of cells of the HO-1+ Mox subtype. Some cells of the M2 and Mox subtypes express bone morphogenic protein-2 (BMP2).

Article Snippet: For IHC, formalin-fixed and paraffin-embedded tissue sections (4 μm in thickness) were blocked with 0.3% H 2 O 2 in methanol at 4°C for 30 min and then subjected to antigen retrieval in trypsin for 30 min at 37°C for anti-CD68 antibody, in citric acid (Antigen Retrieval Solution pH 6; Iatron Laboratories Inc., Tokyo, Japan) for anti-CD 163 antibody, anti-iNOS antibody, anti-HO-1 antibody, and anti-OPN antibody, and in EDTA (pH 9.0; Nichirei, Tokyo, Japan) in an autoclave (2 atmospheres, 121°C, 20 min) for anti-CD206 antibody.

Techniques: