membrane based antibody array  (R&D Systems)

Journal: Heliyon

Article Title: FGF21/FGFR1-β-KL cascade in cardiomyocytes modulates angiogenesis and inflammation under metabolic stress ☆

doi: 10.1016/j.heliyon.2023.e14952

Figure Lengend Snippet: Proteomics-based analysis of candidates of FGFR1-β-KL signaling in myoblasts. A Strategy for cytokine array (medium from C2C12) and MS analysis (cell lysates from H9C2) (created with Biorender.com ). B Log2 fold-change (FC) ( left: HFHG vs Veh) ( right: dual O/E + HFHG vs Veh) of secreted proteins from C2C12 with control/Veh (n = 1) or dual O/E (n = 1). Arrays with duplicate antibody spots and p values were determined by a Z-test (p < 0.05). The dashed line indicates the change to Veh. C log2 transformed mean values to positive control for comparing secreted proteins from HFHG-stimulated C2C12 with or without dual O/E. D-E Pathways database analyses on enriched proteins from D cytokine array and E MS datasets (n = 3/group). Pie chart indicating the arbitrary classification of pathways into inflammation and angiogenesis-related function. F Representation of significantly upregulated and downregulated proteins from identified pathways in E. G-H Volcano dot plots of gene/protein score from targets identified in C and F, from G RNA-seq data from diabetic mouse myocardium (red dots are significantly altered) and H MS datasets from diabetic mouse and human myocardium (n = 3/group). I Bar graph representation of some proteins from H. Bars with green borders were significantly altered compared to their corresponding controls.

Article Snippet: Cytokine Array – Mouse Cytokine Antibody Array (Membrane, L308 Targets) , RayBiotech , Cat# AAM-BLM-1.

Techniques: Transformation Assay, Positive Control, RNA Sequencing Assay

Journal: Heliyon

Article Title: FGF21/FGFR1-β-KL cascade in cardiomyocytes modulates angiogenesis and inflammation under metabolic stress ☆

doi: 10.1016/j.heliyon.2023.e14952

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cytokine Array – Mouse Cytokine Antibody Array (Membrane, L308 Targets) , RayBiotech , Cat# AAM-BLM-1.

Techniques: Cell Culture, Recombinant, Plasmid Preparation, Bradford Assay, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Ab Array, Bicinchoninic Acid Protein Assay, Derivative Assay, Transplantation Assay, Software

Journal: Frontiers in Immunology

Article Title: Dynamic Regulation of the Nexus Between Stress Granules, Roquin, and Regnase-1 Underlies the Molecular Pathogenesis of Warfare Vesicants

doi: 10.3389/fimmu.2021.809365

Figure Lengend Snippet: ISRIB treatment protects against arsenical-mediated inflammation by attenuating Roquin and Regnase-1 degradation in murine skin. (A) Left: representative immunofluorescence imaging of G3BP1-positive SGs (red) in HaCaT cells 3 h after co-treatment with ISRIB (100 nM) and PAO (0.5 µM). Nuclei were stained with DAPI. Right: Quantitative analysis of the percentage of cells expressing SGs in each treatment group (scale bar 50 µm). (B–H) Effects of topical ISRIB (200 µg/mouse) treatment 5-10 min after cutaneous PAO (100 µg/mouse) exposure in Ptch1 +/- /SKH-1 mice. Mice were treated for 6 h before sacrifice. (B) Representative Western blot showing the expression of p-eIF2α, TIA1, and eIF3A after treatment in skin tissue lysates from each treatment group. β-actin was used as the endogenous loading control. (C) Representative immunofluorescence imaging of TIA1 (red) and G3BP1 (green) expression in skin sections from each treatment group. Nuclei were stained with DAPI (scale bar 50 µm). (D) Representative images of H&E-stained skin sections from each group showing the infiltration of inflammatory immune cells (scale bar 50 µm). (E) Left: representative images of skin sections from each treatment group showing the expression of the macrophage marker F4/80, detected by immunohistochemistry. Right: quantitative analysis of the percentage of F4/80 cells per field in each treatment group (scale bar 50 µm). (F) Quantitative analysis of the relative expression of Il1β and Il6 mRNA, assessed by RT-PCR, in skin sections from each treatment group. (G) The relative expression of specific cytokines in skin tissue lysates was assessed by cytokine array. Skin lysates from 4–5 mice from each treatment group were pooled and incubated with the mouse cytokine antibody array membrane. Representative images of the cytokine arrays (top panel) and the histogram depicting densitometry data (bottom panel) are shown. The expression of GCSF (1), IL-6 (2), IL-9 (3), MCP-1 (4), MCP-5 (5), TNFR1 (6) and thrombopoietin (TPO) (7) were significantly higher in PAO-treated skin lysates than in controls but the increased expression was reduced more than 50% in the mice treated with ISRIB after PAO. Cropped array images of these cytokine mediators are presented separately (right panel). Densitometry data were analyzed by excel-based analysis software tools. FC (fold change) (H) Representative Western blots showing Roquin and Regnase-1 expression in skin lysates from each treatment group. β-actin was used as the endogenous loading control. p < 0.05*, p < 0.001***, p < 0.0001**** show significant level.

Article Snippet: Precleared protein (50 µg) lysates of murine skin from at least 5 individual mice from each group were pooled and incubated with a RayBio Mouse Cytokine Antibody Array C3 membrane (CODE: AAM-CYT-3-2) according to the manufacturer’s protocol (RayBiotech, Norcross, GA, USA).

Techniques: Immunofluorescence, Imaging, Staining, Expressing, Western Blot, Marker, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Incubation, Ab Array, Software