Journal: Journal of Neuroscience

Article Title: System xc Activity and Astrocytes Are Necessary for Interleukin-1 -Mediated Hypoxic Neuronal Injury

doi: 10.1523/jneurosci.2459-07.2007

Figure Lengend Snippet: Figure 2. Role of IL-1RI signaling in the potentiation of hypoxic neuronal injury by IL-1 in vitro. A, B, Mixed cortical cell cultures were treated with 1 ng/ml IL-1 for 20–24 h in the presenceorabsenceofrIL-1ra(10–1000ng/ml;A)oranti-IL-1RI(0.1–100g/ml;B),washed, andthendeprivedofoxygen(5h).Thepercentageoftotalneuronalcelldeathwasdetermined 20–24hlater.Anasteriskindicatesvaluessignificantlygreaterthanhypoxiaalone,whereas# denotes a significant diminution of the IL-1-mediated increase in injury (IL-1) as deter- mined by one-way ANOVA followed by a Student–Newman–Keuls t test. Significance was assessed at p 0.05 (n 3–9 cultures pooled from 2–3 different experiments).

Article Snippet: In experiments using recombinant mouse IL-1ra (rIL-1ra) (R & D Systems) or monoclonal anti-mouse IL-1RI antibody directed against the extracellular domain of mouse IL-1RI (MAB7711; R & D Systems), the respective solutions were prepared with IL-1 (1.5 ng/ml; 1.5 ) and added as described above.

Techniques: In Vitro

Journal: Journal of Neuroimmune Pharmacology

Article Title: Heterogeneity of Microglial Activation in the Innate Immune Response in the Brain

doi: 10.1007/s11481-009-9164-4

Figure Lengend Snippet: Genes and antibodies used for profiling of alternative activation and acquired deactivation

Article Snippet: IL1RN , NM_031167 , IL-1 receptor antagonist , WB, ELISA-R&D Systems (AF 480NA).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Brain Interleukin-1β and the Intrinsic Receptor Antagonist Control Peripheral Toll-Like Receptor 3-Mediated Suppression of Spontaneous Activity in Rats

doi: 10.1371/journal.pone.0090950

Figure Lengend Snippet: Poly I:C-induced spontaneous activity in the presence of i.c.v. infusion of rat recombinant IL-1 receptor antagonist (rrIL-1ra, 1 µg/day). Infusion (at a rate of 1 µl/h) was started 1 day before poly I:C injection and continued for 5 days. The line plots represent the percent change in spontaneous activity from baseline. Spontaneous activity was measured from the onset of and throughout the dark period. Arrows depict the time of poly I:C injection. N = 5–8 per experimental group. *P<0.05, versus poly I:C + vehicle infusion group.

Article Snippet: The membrane was preblocked with 5.0% skim milk (WAKO, Tokyo, Japan) in PBS containing 0.1% Tween-20 (PBS-T) at room temperature for 1 h, and then incubated overnight in goat anti-mouse IL-1ra primary antibody (1∶500; R&D Systems, Inc.) at 4°C.

Techniques: Activity Assay, Recombinant, Injection

Journal: PLoS ONE

Article Title: Brain Interleukin-1β and the Intrinsic Receptor Antagonist Control Peripheral Toll-Like Receptor 3-Mediated Suppression of Spontaneous Activity in Rats

doi: 10.1371/journal.pone.0090950

Figure Lengend Snippet: Tissue samples (cerebral cortex, hippocampus, hypothalamus, and cerebellum) were prepared 5 h and 1, 3, 5, and 7 days after poly I:C injection. The bar plots show the levels of IL-1ra mRNA as the fold change relative to the corresponding vehicle-injected group after calibration with the GAPDH mRNA level. N = 4–5 per experimental group. *P<0.05 versus vehicle-injected group.

Article Snippet: The membrane was preblocked with 5.0% skim milk (WAKO, Tokyo, Japan) in PBS containing 0.1% Tween-20 (PBS-T) at room temperature for 1 h, and then incubated overnight in goat anti-mouse IL-1ra primary antibody (1∶500; R&D Systems, Inc.) at 4°C.

Techniques: Injection

Journal: PLoS ONE

Article Title: Brain Interleukin-1β and the Intrinsic Receptor Antagonist Control Peripheral Toll-Like Receptor 3-Mediated Suppression of Spontaneous Activity in Rats

doi: 10.1371/journal.pone.0090950

Figure Lengend Snippet: (A) Immunoblotting result. rrIL-1ra (300, 1000 pg) was detected by cross-reactivity of the anti-mouse neutralizing antibody. Infusion (100 ng/µl, at a rate of 1 µl/h) was started 7 days before poly I:C injection and continued for 5 days. (B) Before injection and (C) after injection of poly I:C or the vehicle. The line plots represent the percent change in spontaneous activity from baseline. Spontaneous activity was measured from the onset of and throughout the dark period. N = 4–8 per experimental group. Arrows depict the time of poly I:C or the vehicle injection. *P<0.05 versus poly I:C + i.c.v. infusion of non-immune IgG group. In this procedure, we injected poly I:C or the vehicle three times (10:00, 18:00, and 10:00 on the next day) to precisely evaluate the effect of neutralizing antibody on the recovery speed of spontaneous activity. We infused antibody from 7 days before poly I:C injection, due to slow diffusion speed of antibody with a big chemical structure.

Article Snippet: The membrane was preblocked with 5.0% skim milk (WAKO, Tokyo, Japan) in PBS containing 0.1% Tween-20 (PBS-T) at room temperature for 1 h, and then incubated overnight in goat anti-mouse IL-1ra primary antibody (1∶500; R&D Systems, Inc.) at 4°C.

Techniques: Western Blot, Injection, Activity Assay, Diffusion-based Assay

Journal:

Article Title: Computational Design and Application of Endogenous Promoters for Transcriptionally Targeted Gene Therapy for Rheumatoid Arthritis

doi: 10.1038/mt.2009.182

Figure Lengend Snippet: Comparison of therapeutic efficacy using Saa3 versus IL1E/IL6P promoter for disease-regulated Il1rn expression. (a) Promoter activity in transduced synovium of naive and arthritic C57Bl/6 mice. Knee joints were injected with 300 ng p24gag equivalent lentivirus encoding PGK, Saa3, or IL1E/IL6P-luciferase. Seven days after transduction, arthritis was induced by intra-articular injection of 180 µg zymosan A. After 24 hours, luciferase activity was assessed ex vivo. Data are represented as individual relative luciferase activities; horizontal bars indicate the means per group. (b) Efficacy of transcriptionally targeted adenoviral vectors expressing Il1rn. NIH-3T3-5xNF-κB-Luc were transduced at a multiplicity of infection (MOI) of 10 with control vector (del) or adenovirus encoding CMV, Saa3, and IL1E/IL6P-driven Il1rn. After 24 hours, cells were transduced at an MOI of 10 with control vector (del) or Ad5.CMV-Il1b. The day thereafter, IL-1β-induced NF-κB activation was assessed by luciferase assay. Data are represented as relative luciferase activities ± SEM (n = 4), and differences were determined using analysis of variance with Dunnett's post-test. **P < 0.01.

Article Snippet: White high-binding flat bottom 96-well plates (Greiner Bio-One, Alphen a/d Rijn, the Netherlands) were coated with the capture antibody rat anti-murine Il1rn (MAB480; R&D Systems, Minneapolis, MN) at 3 μg/ml in 0.1 mol/l carbonate buffer pH 9.6 and incubated overnight at 4 °C.

Techniques: Comparison, Drug discovery, Expressing, Activity Assay, Injection, Luciferase, Transduction, Ex Vivo, Infection, Control, Plasmid Preparation, Activation Assay