Journal: Advanced healthcare materials

Article Title: Biomimetic Nanocarriers of Pro-Resolving Lipid Mediators for Resolution of Inflammation in Atherosclerosis.

doi: 10.1002/adhm.202302238

Figure Lengend Snippet: Figure 5. Flow cytometry analysis of the expression of SPMs receptors (ChemR23, GPR18, FPR2, and Lgr6) on the surface of A) EC (bEnd.3 cells), B) SMC (MOVAS cells), and C) macrophages (RAW 264.7 cells). Above is an overlay of representative histograms showing the expression of SPMs receptors on quiescent or activated cells (green and pink lines, respectively), and the respective isotype (red line). Below is a graphical representation of the percentage of the specified SPM receptor-positive cells. The data are expressed as mean ± S.D. from three independent experiments carried out in duplicate. *p < 0.05, **p < 0.01, and ****p < 0.0001; comparisons were made between quiescent and activated cells. Statistical variance and significance were calculated using one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: The pellet obtained was washed and reconstituted in 100 μL of PBS containing 0.5% PFA, 5 mm EDTA, 5% FBS (FACS buffer), and the fluorochromecoupled antibody at a concentration of 5 μg mL−1: mouse anti-ChemR23 PE-conjugated Antibody (R&D Systems, cat. no. FAB7610P), rabbit antiGPR18 FITC-conjugated Antibody (Novus Biologicals, cat. no. NBP224918F), rabbit anti-FPR2 FITC-conjugated Antibody (Novus Biologicals, cat. no. NLS1878F), and rabbit anti-Lfr6 FITC-conjugated Antibody (Novus Biologicals, cat. no. FAB8458RF).

Techniques: Flow Cytometry, Expressing

Journal: Advanced Healthcare Materials

Article Title: Biomimetic Nanocarriers of Pro‐Resolving Lipid Mediators for Resolution of Inflammation in Atherosclerosis

doi: 10.1002/adhm.202302238

Figure Lengend Snippet: Flow cytometry analysis of the expression of SPMs receptors (ChemR23, GPR18, FPR2, and Lgr6) on the surface of A) EC (bEnd.3 cells), B) SMC (MOVAS cells), and C) macrophages (RAW 264.7 cells). Above is an overlay of representative histograms showing the expression of SPMs receptors on quiescent or activated cells (green and pink lines, respectively), and the respective isotype (red line). Below is a graphical representation of the percentage of the specified SPM receptor‐positive cells. The data are expressed as mean ± S.D. from three independent experiments carried out in duplicate. * p < 0.05, ** p < 0.01, and **** p < 0.0001; comparisons were made between quiescent and activated cells. Statistical variance and significance were calculated using one‐way ANOVA with Tukey's post‐hoc test.

Article Snippet: The pellet obtained was washed and reconstituted in 100 µL of PBS containing 0.5% PFA, 5 m m EDTA, 5% FBS (FACS buffer), and the fluorochrome‐coupled antibody at a concentration of 5 µg mL −1 : mouse anti‐ChemR23 PE‐conjugated Antibody (R&D Systems, cat. no. FAB7610P), rabbit anti‐GPR18 FITC‐conjugated Antibody (Novus Biologicals, cat. no. NBP2‐24918F), rabbit anti‐FPR2 FITC‐conjugated Antibody (Novus Biologicals, cat. no. NLS1878F), and rabbit anti‐Lfr6 FITC‐conjugated Antibody (Novus Biologicals, cat. no. FAB8458RF).

Techniques: Flow Cytometry, Expressing

Journal: Arthritis Research & Therapy

Article Title: Chemerin activates fibroblast-like synoviocytes in patients with rheumatoid arthritis

doi: 10.1186/ar3475

Figure Lengend Snippet: Expression of ChemR23 in rheumatoid arthritis and osteoarthritis synovial tissue samples . ChemR23 expression in synovial tissue samples examined by immunohistochemistry that were obtained from five rheumatoid arthritis (RA) patients (A) and three osteoarthritis (OA) patients (B) . All sections were counterstained with hematoxylin. Sections of RA synovial tissues were also double-stained with CD68, CD1a, DC-LAMP or vimentin, and ChemR23 and were analyzed by fluorescence microscopy as follows: (C) CD68, (D) ChemR23, (E) merged image of (C) and (D), (F) CD1a, (G) ChemR23, (H) merged image of (F) and (G), (I) DC-LAMP, (J) ChemR23, (K) merged image of (I) and (J), (L) vimentin, (M) ChemR23 and (N) merged image of (L) and (M). Arrows indicate double-positive cells. Original magnification ×100 in (A) and (B) and ×400 in (C) through (N). Whole-mount specimens obtained from RA patients were stained with anti-ChemR23 pAb (green) or phalloidin (blue) and then analyzed by confocal laser-scanning microscopy. (O) x - y view of synovial tissue sectioning. x - z image (P) and y-z image (Q) of synovial tissue sectioning are displayed below and to the right of the x - y view in (O). Scale bars = 80 μm. White straight line boxes indicate identical cell in each direction (O-Q). Dashed line connects the boxes.

Article Snippet: Serial sections were then incubated for two hours at 4°C with 1 μg/ml rabbit anti-chemerin pAb (affinity-purified from rabbit serum immunized with glutathione S -transferase-chemerin (E 21 -S 157 ) fusion protein; provided by KAN Research Institute, Kobe, Japan), 1 μg/ml rabbit anti-ChemR23 pAb (Acris Antibodies, Herford, Germany) or normal rabbit immunoglobulin G (IgG) (Sigma-Aldrich, St Louis, MO, USA) as an isotype control.

Techniques: Expressing, Immunohistochemistry, Staining, Fluorescence, Microscopy, Confocal Laser Scanning Microscopy

Journal: Arthritis Research & Therapy

Article Title: Chemerin activates fibroblast-like synoviocytes in patients with rheumatoid arthritis

doi: 10.1186/ar3475

Figure Lengend Snippet: Expression of chemerin and ChemR23 in rheumatoid arthritis and osteoarthritis synovial tissue samples . (A) Western blots of chemerin protein expression in three rheumatoid arthritis (RA) and four osteoarthritis (OA) synovial tissues. M, protein molecular weight marker. (B) Relative protein expression of chemerin to β-actin in RA and OA synovial tissues. * P < 0.05 relative to RA synovial tissue. (C) Western blots of ChemR23 protein expression in three RA and four OA synovial tissues. M, protein molecular weight marker. (D) Relative expression of ChemR23 protein to β-actin. * P < 0.05 relative to RA synovial tissue.

Article Snippet: Serial sections were then incubated for two hours at 4°C with 1 μg/ml rabbit anti-chemerin pAb (affinity-purified from rabbit serum immunized with glutathione S -transferase-chemerin (E 21 -S 157 ) fusion protein; provided by KAN Research Institute, Kobe, Japan), 1 μg/ml rabbit anti-ChemR23 pAb (Acris Antibodies, Herford, Germany) or normal rabbit immunoglobulin G (IgG) (Sigma-Aldrich, St Louis, MO, USA) as an isotype control.

Techniques: Expressing, Western Blot, Molecular Weight, Marker

Journal: Arthritis Research & Therapy

Article Title: Chemerin activates fibroblast-like synoviocytes in patients with rheumatoid arthritis

doi: 10.1186/ar3475

Figure Lengend Snippet: Expression of chemerin and ChemR23 by rheumatoid arthritis fibroblast-like synoviocytes . (A) and (B) Chemerin expression stimulated by TNF-α (A) or IFN-γ (B) and evaluated by ELISA using supernatants of cultured fibroblast-like synoviocytes (FLSs) isolated from synovial tissue samples taken from rheumatoid arthritis (RA) patients ( n = 4). FLSs (2 × 10 4 cells/well) were stimulated at 37°C for 48 hours with TNF-α (0.1, 1 or 10 ng/ml) (A) or IFN-γ (1, 10 or 100 ng/ml) (A). Data in (A) and (B) are presented as means (± SEM) of one of four independent experiments analyzed in triplicate. * P < 0.05 relative to control. (C) through (E) Immunocytochemistry showing double-staining of cultured RA FLSs with (C) vimentin (D) ChemR23 and (E) a merged image of (C) and (D). (F) Western blot of ChemR23 protein expression in RA FLSs following TNF-α (10 ng/ml), IFN-γ (100 ng/ml), transforming growth factor (TGF)-β1 (1 ng/ml), IL-1β (5 ng/ml) or IL-6 (20 ng/ml) stimulation at 37°C for 24 hours.

Article Snippet: Serial sections were then incubated for two hours at 4°C with 1 μg/ml rabbit anti-chemerin pAb (affinity-purified from rabbit serum immunized with glutathione S -transferase-chemerin (E 21 -S 157 ) fusion protein; provided by KAN Research Institute, Kobe, Japan), 1 μg/ml rabbit anti-ChemR23 pAb (Acris Antibodies, Herford, Germany) or normal rabbit immunoglobulin G (IgG) (Sigma-Aldrich, St Louis, MO, USA) as an isotype control.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Isolation, Immunocytochemistry, Double Staining, Western Blot

Journal: Advanced Healthcare Materials

Article Title: Biomimetic Nanocarriers of Pro‐Resolving Lipid Mediators for Resolution of Inflammation in Atherosclerosis

doi: 10.1002/adhm.202302238

Figure Lengend Snippet: Flow cytometry analysis of the expression of SPMs receptors (ChemR23, GPR18, FPR2, and Lgr6) on the surface of A) EC (bEnd.3 cells), B) SMC (MOVAS cells), and C) macrophages (RAW 264.7 cells). Above is an overlay of representative histograms showing the expression of SPMs receptors on quiescent or activated cells (green and pink lines, respectively), and the respective isotype (red line). Below is a graphical representation of the percentage of the specified SPM receptor‐positive cells. The data are expressed as mean ± S.D. from three independent experiments carried out in duplicate. * p < 0.05, ** p < 0.01, and **** p < 0.0001; comparisons were made between quiescent and activated cells. Statistical variance and significance were calculated using one‐way ANOVA with Tukey's post‐hoc test.

Article Snippet: Corresponding isotypes were used as controls: rat anti‐IgG2B PE‐conjugated Antibody (for ChemR23 receptor; R&D Systems, cat. no. IC013P), and rabbit anti‐IgG FITC‐conjugated Antibody (for GPR18 and FPR2 receptors; R&D Systems, cat. no. IC1051F) at a concentration of 10 µL/10 6 cells), as well as mouse anti‐IgG F(ab) FITC‐conjugated Antibody (for the Lgr6 receptor; Novus Biologicals, cat. no. NBP1‐97143) at a concentration of 5 µg mL −1 .

Techniques: Flow Cytometry, Expressing

Journal: Annals of diagnostic pathology

Article Title: Specialized pro-resolving receptors are expressed in salivary glands with Sjögren’s syndrome

doi: 10.1016/j.anndiagpath.2021.151865

Figure Lengend Snippet: Primary antibodies used in this study.

Article Snippet: Moreover, rabbit anti-GPR32, rabbit anti-BLT1, rabbit anti-CMKLR1, Alexa fluor 488 conjugated anti-rabbit IgG, phosphate buffered saline (PBS), 4′,6-diamidino-2-phenylindole (DAPI), triton X-100, sodium citrate, xylene and ethanol were purchased from Thermo Fisher Scientific (Waltham, MA).

Techniques: