rat cytokine antibody arrays  (R&D Systems)

Journal: International Journal of Medical Sciences

Article Title: Hyperbaric oxygen protects against periodontal bone loss by modulating inflammation and bone remodeling via RANKL/OPG expression in ligature-induced periodontitis

doi: 10.7150/ijms.122857

Figure Lengend Snippet: HBOT reduces gingival cytokine and chemokine expression in ligature-induced periodontitis. (A) Representative cytokine array blots showing expression profiles of 10 key inflammatory mediators (red boxes) in gingival tissues collected at day 14 and day 28 from Sham, PD+RECOV (natural recovery), PD (periodontitis only), PD+EHBOT (early HBOT), and PD+LHBOT (late HBOT) groups. The targeted proteins included: 1. CINC-1 (CXCL1), 2. CINC-2α/β (CXCL3), 3. sICAM-1, 4. IL-1α, 5. IL-1β, 6. IL-1 receptor antagonist (IL-1ra), 7. LIX (CXCL5), 8. L-selectin (CD62L), 9. Thymus chemokine (CCL25), and 10. TIMP-1. (B) Quantitative analysis of cytokine and chemokine expression at day 14. Periodontitis induced marked increases in several pro-inflammatory cytokines and chemokines. Early HBOT significantly suppressed most inflammatory mediators compared to the PD and PD+RECOV groups. (C) Quantitative analysis at day 28. PD-induced cytokine elevation persisted, while both early and late HBOT treatments effectively reduced IL-1α, IL-1β, IL-1ra, sICAM-1, CINC-1, CINC-2α/β, LIX, and thymus chemokine levels. Notably, early HBOT more effectively reduced thymus chemokine expression than natural recovery. Data are presented as mean ± SD. n = 6 per group. Panel 2A shows a cytokine dot-array membrane (R&D Systems ARY008) from a single, intact exposure; the published panel was border-cropped only to remove blank margins (global linear contrast, no compositing). See Supplementary for the full, uncropped membrane and original X-film/RAW.

Article Snippet: Protein extracts (200 μg per sample) were applied to Rat Cytokine Antibody Arrays (ARY008, R&D Systems, Inc., USA), which detect a panel of 34 inflammatory cytokines, chemokines, and adhesion molecules.

Techniques: Expressing, Membrane

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Biostimulatory Micro-fragmented Nanofiber-Hydrogel Composite Improves Mesenchymal Stem Cell Delivery and Soft Tissue Remodelling

doi: 10.1002/smll.202202309

Figure Lengend Snippet: (A) Representative immunofluorescent staining images showing the infiltrated endothelial cells (stained with RECA-1 in red and alpha smooth muscle actin, αSMA. in green) inside the matrices at POD 14 and 28. Scale bar: 100 μm. (B) Quantitative data presenting the infiltrated endothelial cells density in the matrix over 28 days (n = 6). (C) Immunofluorescent staining images showing the adipogenesis inside the matrix at POD 14 and 28; perilipin-1 was stained in green, and adiponectin-1 stained with Acrp-30 in red. Scale bar: 200 μm. (D) Profiling of immunomodulation (left) and tissue remodelling (right) cytokine proteins production in mfNHC and mfNHC-MSC matrix at POD 14 and 28. Statistical significance was calculated by two-tailed Student t-test. N.S.P>0.05, **P<0.01. Data are presented as Mean ± Standard Error of the Mean.

Article Snippet: To characterize the inflammation- and regeneration-related cytokines and chemokines mediated by the injected mfNHC and/or MSCs, a rat cytokine antibody proteome profiler array (ARY030, R&D Systems) was used to analyse the tissue samples collected at POD 14 and 28.

Techniques: Staining, Two Tailed Test

Journal: Autophagy

Article Title: An autophagic deficit in the uterine vessel microenvironment provokes hyperpermeability through deregulated VEGFA, NOS1, and CTNNB1.

doi: 10.1080/15548627.2020.1778292

Figure Lengend Snippet: Figure 3. Characterization of cellular abnormalities in OVX atg7d/d uteri. (A) Immunofluorescence staining of SQSTM1 and PECAM1 in OVX atg7d/d uteri. Localization of SQSTM1 (green signal) in vascular mural cell layers surrounding PECAM1+ (red signal) endothelial cells. The area marked with a yellow rectangle is enlarged in the right panel. Myo, myometrium; le, luminal epithelium; s, stroma; VSMC, vascular smooth muscle cell; EC, endothelial cells. Scale bar: 100 μm. (B) The average number of USCs per mouse harvested by enzymatic digestion method. Usually 3–5 mice were used for USC preparation for cell sorting, culture, RNA preparation, or proteome array experiments. 1*p = 0.029, t = 1.950, df = 41; 2*p = 0.0271, t = 2.181, df = 10. (C) Co-immunofluorescence staining of cell markers and SQSTM1 in USCs isolated from E2-injected OVX Atg7 f/f and atg7d/d mice. Red puncta in subsets of atg7d/d USCs indicating the sites of SQSTM1 accumulation. Scale bar: 20 μm. (D) USCs were isolated from E2-injected OVX Atg7 f/f and atg7d/d mice and subjected to cell sorting. Five sets of independent analyses were performed using pooled samples from 3 or 4 mice and a representative set is shown here. (E) Co-immunofluorescence staining of MCAM (green signal) and other markers in USCs isolated from E2-injected OVX atg7d/d mice. Subsets of MCAM+ cells also expressed DES, CSPG4, or ACTA2. White scale bar: 20 μm.(F) Immunofluorescence staining of COL1A1 and PECAM1 in OVX Atg7 f/f and atg7d/d uteri. Enlarged images of subepithelial stromal regions are shown. Scale bar: 100 (left panel) or 10 μm (middle and right panels).

Article Snippet: Semiquantitative cytokine detection was performed using Proteome ProfilerTM Antibody Arrays for 111 different analytes (R&D Systems, ARY028) and Proteome ProfileTM Mouse Phospho-RTK Arrays (R&D Systems, ARY014).

Techniques: Immunofluorescence, Staining, FACS, Isolation, Injection

Journal: Autophagy

Article Title: An autophagic deficit in the uterine vessel microenvironment provokes hyperpermeability through deregulated VEGFA, NOS1, and CTNNB1.

doi: 10.1080/15548627.2020.1778292

Figure Lengend Snippet: Figure 5. VEGFA- and NO-regulated hyperpermeability in atg7d/d uteri. (A) USC lysates were prepared from 5 or 6 OVX Atg7 f/f and atg7d/d mice and assayed with the Mouse XL Cytokine Array containing 111 secreted factors. Three independent experiments (17 mice per group) were performed and factors showing more than 1.2-fold increase in atg7d/d USC lysates in all three sets are shown in Table 1. (B) USC lysates were prepared from 5 or 6 OVX Atg7 f/f and atg7d/d mice and assayed with the Proteome ProfileTM Mouse Phospho-Receptor Tyrosine Kinase (RTK) Array. Three independent experiments (17 mice per group) were performed and factors showing more than 1.2-fold increase in atg7d/d USC lysates in all three sets are shown in Table 2. (C) qPCR analysis of factors with significant changes in both 3ʹ mRNA sequencing and cytokine array. Relative gene expression was normalized to ribosomal protein L7 (Rpl7). Values represent the mean ± S.E.M. 1*p = 0.0141, t = 2.564, df = 10; 2***p < 0.0001, t = 7.918, df = 10; 3***p < 0.0001, t = 6.207, df = 10; 4***p < 0.0001, t = 17.43, df = 10; 5*p = 0.0436, t = 1.895, df = 10; 6***p < 0.0001, t = 13.26, df = 10. (D) Age-dependent increase of VEGFA and NOS1 levels in atg7d/d uteri. Uteri were collected from random cycling mice. 4 w, 4-week-old; 8 w, 8-week-old; 1 y, > 1-year-old. Each sample was prepared from a single mouse. EDN1 levels did not show an age-dependent increase like VEGFA and NOS1. (E) Experimental scheme for examining the effect of L-NAME on plasma extravasation in OVX atg7d/d uteri. Mice received daily intraperitoneal injections of L-NAME (0.625 mg/0.1 ml PBS) for 10 (Miles assay) or 14 d (western blotting). Uteri were collected 24 h after the last injection and subjected to further analyses. (F) Uterine Evans blue content after L-NAME injection measured by Miles assay. 1**p = 0.0049, t = 2.934, df = 16; 2ns, p = 0.0506, t = 1.739, df = 16; 3*p = 0.047, t = 1.780, df = 15. (G) Western blot analyses of CTNNB1 in L-NAME-injected atg7d/d uteri. 1ns, p = 0.1510, t = 1.129, df = 6; 2*p = 0.0176, t = 2.708, df = 6. (H) Co- immunofluorescence staining of CTNNB1 (green) and PECAM1 (red) in OVX atg7d/d uteri. Scale bar: 50 μm. White arrows indicate CTNNB1 signal in PECAM1-positive endothelial cells.

Article Snippet: Semiquantitative cytokine detection was performed using Proteome ProfilerTM Antibody Arrays for 111 different analytes (R&D Systems, ARY028) and Proteome ProfileTM Mouse Phospho-RTK Arrays (R&D Systems, ARY014).

Techniques: Sequencing, Gene Expression, Clinical Proteomics, Western Blot, Injection, Immunofluorescence, Staining

Journal: Glia

Article Title: Elevated fibroblast growth factor-inducible 14 expression transforms proneural-like gliomas into more aggressive and lethal brain cancer

doi: 10.1002/glia.24018

Figure Lengend Snippet: Protein profiling summary. The top 10 proteins upregulated in Fn14-low and Fn14-high rat tumors compared to normal rat brain as determined using R&D Systems Rat XL Cytokine Array Kit are listed. Proteins that are specifically overexpressed in the Fn14-high tumors compared to both normal brain and Fn14-low tumors are written in bold type

Article Snippet: Protein profiling was performed using the Rat XL Cytokine Antibody Array Kit (R&D Systems ARY030) according to manufacturer's instructions.

Techniques: