Journal: Mediators of Inflammation

Article Title: Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner

doi: 10.1155/2022/9946439

Figure Lengend Snippet: The effect of CL on the expression of GFAP (a) and secretion of IL-1 β (b) by human U118 MG astrocytic cells. CL (20 μ g/ml) or its vehicle solution was added to U118 MG cell cultures 15 min before exposure to LPS (400 ng/ml) or the vehicle solution. After 48 h, immunoblotting was performed using cell lysates (a), and IL-1 β concentration in cell supernatants was measured by an ELISA (b). Data (means ± SEM) are expressed as fold change in protein levels compared to control cells exposed to growth medium only and are normalized to β -actin (a). The detection limit of the ELISA is presented as the dotted line (b). ∗ P < 0.05 and ∗∗ P < 0.01 according to Tukey's post-hoc test. P and F values for the one-way randomized blocks ANOVA are shown. Representative bands of immunostained GFAP and β -actin from each treatment group are shown (c). Uncropped immunoblot images are provided as Supplementary Figure .

Article Snippet: Concentrations of MCP-1, IP-10, and IL-1 β in supernatants were measured 48 h later using PeproTech ELISA development kits according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Mediators of Inflammation

Article Title: Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner

doi: 10.1155/2022/9946439

Figure Lengend Snippet: The effect of CL on human U118 MG astrocytic cell MCP-1 secretion (a)–(c) and their viability (d) in serum-containing medium. Increasing concentrations of CL or its vehicle solution was added to U118 MG astrocytic cell cultures (a, d), or CL was added 15 min before exposure to LPS (400 ng/ml) (b) or a combination of IFN- γ (400 U/ml) plus IL-1 β (20 pg/ml) (c). After 48 h, concentrations of MCP-1 in cell supernatants were measured by an ELISA, and the viability of U118 MG astrocytic cells was measured by the MTT assay. Data are presented as means ± SEM. Cell viability in (d) is normalized to values obtained from U118 MG cultures not exposed to CL or its vehicle solution. ∗∗ P < 0.01 according to Dunnett's post-hoc test. P and F values for the one-way randomized blocks ANOVA are shown. The detection limit of the ELISA is presented as the dotted line.

Article Snippet: Concentrations of MCP-1, IP-10, and IL-1 β in supernatants were measured 48 h later using PeproTech ELISA development kits according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, MTT Assay

Journal: Mediators of Inflammation

Article Title: Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner

doi: 10.1155/2022/9946439

Figure Lengend Snippet: The effect of CL on human U118 MG astrocytic cell MCP-1 secretion in serum-free medium. Increasing concentrations of CL or its vehicle solution was added to U118 MG astrocytic cell cultures (a), or CL was added 15 min before exposure to LPS (400 ng/ml) (b), a combination of IFN- γ (400 U/ml) plus IL-1 β (20 pg/ml) (c). After 48 h, the concentrations of MCP-1 in cell supernatants were measured by an ELISA. Data are presented as means ± SEM. ∗ P < 0.05 and ∗∗ P < 0.01 according to Dunnett's post-hoc test. P and F values for the one-way randomized blocks ANOVA are shown. The detection limit of the ELISA is presented as the dotted line.

Article Snippet: Concentrations of MCP-1, IP-10, and IL-1 β in supernatants were measured 48 h later using PeproTech ELISA development kits according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Adenosine deaminase modulates metabolic remodeling and orchestrates joint destruction in rheumatoid arthritis

doi: 10.1038/s41598-021-94607-5

Figure Lengend Snippet: Showing ADA Activity and Cytokine levels in Rheumatoid arthritis patients. ( a ) ADA activity in RA patients representing group with Low ADA and High ADA when compared to healthy controls. ( b ) ADA activity of two isoenzymes ADA 1 and ADA 2 (*representing a p value of ≤ 0.05) ( c ) Levels of Cytokines in the serum of RA patients. ( d ) One-way ANOVA analysis of Cytokines and ADA levels in healthy controls, Low ADA and High ADA RA patients. (p-value of ≤ 0.05 is considered significant). All the plots were made using Origin 2020.

Article Snippet: TNFα (Cat. No. CHC1753), TGFβ (Cat. No. CHC1683), IFNγ (Cat. No. CHC1233), and IL-10 (Cat. No. CHC1323) (Invitrogen Cytoset Antibody Pair Kit , California, USA) and IL-6 (Cat. No. 900 M16) and sRANKL (Cat. No. 900 M142) (Peprotech Mini ABTS ELISA Kit, New Jersey, USA) were estimated using manufacturers’ protocols and calculated by the 4-parameter curve-fit method.

Techniques: Activity Assay

Journal: Scientific Reports

Article Title: Adenosine deaminase modulates metabolic remodeling and orchestrates joint destruction in rheumatoid arthritis

doi: 10.1038/s41598-021-94607-5

Figure Lengend Snippet: ADA is involved in joint remodeling in rheumatoid arthritis—in vitro studies to investigate the role of ADA in joint remodeling in the affected joint. ( a ) Increased proliferation of Synoviocytes, ( b ) Decreased cell viability in primary chondrocytes. ( c ) Increased Osteoclast differentiation of macrophages and ( d ) qPCR analysis showing the expression of osteoclast markers after 72 h exposure to ADA. ( e ) Increased cytokine levels in the spent medium after 72 exposure to ADA. ( f ) P2Y 2 , P2Y 4 , and P2Y 6 receptor expression in macrophages after 72 h of ADA exposure. (showing *representing a p value of ≤ 0.05, and **representing a p value of ≤ 0.005). ( g ) Increased ROS in macrophages after ADA exposure (p values of ADA to CTRL on all time points are ≤ 0.005). All the plots were made using Origin 2020.

Article Snippet: TNFα (Cat. No. CHC1753), TGFβ (Cat. No. CHC1683), IFNγ (Cat. No. CHC1233), and IL-10 (Cat. No. CHC1323) (Invitrogen Cytoset Antibody Pair Kit , California, USA) and IL-6 (Cat. No. 900 M16) and sRANKL (Cat. No. 900 M142) (Peprotech Mini ABTS ELISA Kit, New Jersey, USA) were estimated using manufacturers’ protocols and calculated by the 4-parameter curve-fit method.

Techniques: In Vitro, Expressing

Journal: ACS Omega

Article Title: Auxiliary Biomembranes as a Directional Delivery System To Control Biological Events in Cell-Laden Tissue-Engineering Scaffolds

doi: 10.1021/acsomega.6b00502

Figure Lengend Snippet: Release of IL-4 and VEGF from gelatin/HA-tyr films. (A, B) Schematic representation of different loading ways of growth factors inside gelatin/HA-tyr cross-linked films. Release profile determined by the ELISA test over 144 h for VEGF (C, D) and IL-4 (E, F) at 37 °C in PBS. For C–F, fresh supernatant was added after each record and three samples for each condition were used.

Article Snippet: The levels of cytokines were measured with ELISA Development Kits (VEGF, IL-4, IL1-RA, IL-6, IL-8, TNF-α, CCL18, PeproTech).

Techniques: Enzyme-linked Immunosorbent Assay