Journal: Journal of Neuroinflammation

Article Title: Laquinimod attenuates inflammation by modulating macrophage functions in traumatic brain injury mouse model

doi: 10.1186/s12974-018-1075-y

Figure Lengend Snippet: Inflammatory responses are suppressed by laquinimod in monocytes following TBI. a Gene expression of inflammatory-related molecules in peripherally derived monocytes as measured by MG468 chip. b–c qPCR validation of iNOS ( b ) and IL-6 ( c ) in peripherally derived monocytes. We studied five to seven mice per group from at least three independent experiments. Bars show mean ± s.e.m. ( n = 5)

Article Snippet: Primers and probes for IL − 6 (Taqman Gene Expression Assay ID Mm00446190) and iNOS (Mm00440502) were purchased from Applied Biosystems. mRNA levels were normalized relative to GAPDH (Applied Biosystems, 4351309), by the formula 2^(−ΔCt), where ΔCt = CtmiR-X-CtGAPDH.

Techniques: Expressing, Derivative Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Mouse Assay

Journal: Physiological Reports

Article Title: Acute administration of interleukin‐6 does not increase secretion of glucagon‐like peptide‐1 in mice. Acute administration of interleukin‐6 does not increase secretion of glucagon‐like peptide‐1 in mice

doi: 10.14814/phy2.13788

Figure Lengend Snippet: Lack of effect of IL‐6 on GLP‐1 Secretion in the Perfused Mouse Small Intestine and in GLP‐1 producing GLUTag cells. (A) GLP‐1 secretion (pmol/L) in perfused mouse small intestines ( n = 6). Infusion of IL‐6 (100 ng/mL) (10–20 min), followed by a period in the absence of IL‐6 (21–39 min), and finally, a period in the presence of 10 nmol/L of bombesin (positive control; 40–45 min). (B) Hormone output (secretion) calculated as area under the curve, with (gray) or without (white) IL‐6 (100 ng/mL), was calculated for each mouse and illustrated collectively as bar and whiskers (Tukey distribution). (C) IL‐6 bioactivity in perfusion samples (#1‐#4) was determined by the ability of these perfusates to induce STAT3 phosphorylation (P‐STAT3) as analyzed by SDS‐PAGE and Western blotting. One‐hundred ng/mL of mIL‐6 was used as positive control (pos.) and Krebs‐Ringer bicarbonate buffer as negative control (neg.). Total STAT3 and α ‐tubulin were used as loading controls. (D) GLP‐1 levels in cell media after 2 h incubation with buffer, 100 or 1000 ng/mL IL‐6, or 10 mmol/L glucose ( n = 4). IL‐6 had no effect on GLP‐1 secretion compared to basal levels in either of these experimental models. Data in panel A and D are shown as mean ± SEM and in panel B as box and whisker (Tukey distribution).

Article Snippet: Test substances consisted of mouse IL‐6 (100 and 1000 ng/mL, R & D Systems, Minneapolis, USA, cat.no.

Techniques: Positive Control, SDS Page, Western Blot, Negative Control, Incubation, Whisker Assay

Journal: Nature Communications

Article Title: IL-6 receptor blockade corrects defects of XIAP-deficient regulatory T cells

doi: 10.1038/s41467-018-02862-4

Figure Lengend Snippet: Anti-IL-6R reduces the expression of IFN-γ in activated Xiap −/− Treg cells. a , b Anti-IL-6R decreases IFN-γ expression in re-stimulated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated with anti-CD3/CD28 and IL-2, or with additional IL-12 as indicated, in the absence or presence of anti-IL-6R (50 μg ml −1 ) for 4 days. iTreg cells were re-stimulated with TPA/A23187 for 24 h and IFN-γ production was determined by ELISA ( a ), or reactivated with TPA/A23187 for 5 h and the expressions of Foxp3 and IFN-γ were determined by intracellular staining ( b ). c , d Anti-IL-6R inhibits IFN-γ expression in human Treg cells. Control and human XIAP-knockdown iTreg cells were stimulated as in ( a , b ), with additional IL-6 as indicated, and secretion ( c ) or intracellular expression ( d ) of IFN-γ was determined. e Inability of anti-TNF or anti-IL-1R to inhibit the production of IFN-γ in activated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated, as described in ( a ), in the presence or absence of anti-TNF or anti-IL-1R (50 μg ml −1 each) for 4 days. Treg cells were re-stimulated with TPA/A23187 for 24 h and the secreted IFN-γ was determined by ELISA. f Anti-IL-6R rescues the impaired suppressive activity of Xiap −/− tTreg cells in vivo. CD45.2 + WT or Xiap −/− tTreg cells were co-transferred with CD45.1 + CD4 + CD25 - effector T cells into male CD45.1 + RagI −/− mice. Anti-IL-6R antibody (500 μg per mouse) was intraperitoneally administrated at day 0, followed by weekly dosing of 500 μg. The body weights of mice were monitored at the indicated time-points. *** P

Article Snippet: Recombinant mouse IL-2, human IL-2 and mouse IL-6 were purchased from eBioScience.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, In Vivo, Mouse Assay