Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 1. Characterization of floxed il-1rn mice. IL-1Ra levels were measured by ELISA in the sera (A) and in spleen (B), liver (C), lung (D), and skin (E) extracts, 4 h after LPS (2 mg/kg) or saline solution (NaCl) injection, in control mice (WT, n = 3) and heterozygous (IL-1Raflx/+, n = 3) and homozygous (IL-1Raflx/flx, n = 3) floxed il-1rn mice. The results represent the mean 6 SEM of IL-1Ra levels. pp , 0.05; ppp , 0.01; pppp , 0.001, versus NaCl-injected mice by ANOVA, two-tailed t test.

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Saline, Injection, Control, Two Tailed Test

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 2. Characterization of conditional hepatocyte-specific IL-1Ra– deficient mice. A, Alb-Cre–mediated deletion of il-1rn exon 2. Exons are depicted as boxes and numbered. The vector-derived sequences and the mouse il-1rn genomic sequences are represented by bold and thin lines, respectively. LoxP sites are shown as black triangles, and FRT sites are shown as double gray triangles. Diagram is not depicted to scale. B, Il-1rn exon 2 copies were quantified in mouse primary hepatocytes and skin by quantitative PCR on genomic DNA. The number of copies of il-1rn exon 2 was normalized to that of il-1rn exon 3. Location of PCR primers are indicated in A (arrows a and b for exon 2, arrows c and d around exon 3). Results represent exon2/exon3 ratio 3 100. Data are mean 6 SEM for hepatocyte-specific IL-1Ra–deficient mice (IL-1RaDH; n = 3) and control mice (WT; n = 3). ppp p , 0.001, by Student t test. C, Production of IL-1Ra by mouse primary hepatocytes. Hepatocytes were cultured in the presence of LPS (1 mg/ml) or with a combination of IL-1b (1 ng/ml) and IL-6 (10 ng/ml). After 48 h, supernatants were collected, and IL-1Ra levels were measured by ELISA. Values represent the mean 6 SEM of IL-1Ra production by cells isolated from hepatocyte-specific IL-1Ra–deficient mice (IL-1RaDH, n = 3) and control mice (WT, n = 3). pppp , 0.001, versus cells isolated from WT mice with similar stimulation assessed by ANOVA, two-tailed t test. ic, intracellular; s, secreted.

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Plasmid Preparation, Derivative Assay, Genomic Sequencing, Real-time Polymerase Chain Reaction, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Isolation, Two Tailed Test

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 3. Contribution of hepatocytes and myeloid cells as sources of circulating IL-1Ra after systemic challenge with LPS. IL-1Ra levels were quantified by ELISA in sera of control mice (WT), IL-1RaDH mice, IL-1RaDM mice, and IL-1RaDH+M mice 4 h (A) or 18 h (B) after 10 mg/kg LPS injection. A and B are two independent experiments. Results are shown as individual values for each mouse (symbols) and mean values (lines). pp , 0.05; ppp , 0.01; pppp , 0.001, versus control mice by ANOVA, two-tailed t test.

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Injection, Two Tailed Test

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 4. Contribution of hepatocytes and myeloid cells as sources of IL-1Ra in the liver and spleen in response to LPS. A and C, Quan- tification of IL-1Ra levels in liver and spleen after LPS challenge. IL-1Ra levels were deter- mined by ELISA in liver (A) or spleen (C) extracts of control mice (WT), IL-1RaDH

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 5. Functional role of hepatocyte- and/or myeloid cell-derived IL-1Ra in the control of LPS-induced lethality. Control mice (WT; n = 19), IL-1RaDH mice (n = 16), IL-1RaDM mice (n = 10), IL-1RaDH+M mice (n = 9), and IL-1Ra–deficient mice (n = 12) were injected i.p. with 10 mg/kg LPS. Survival was scored for 6 d. pp , 0.05 versus WT mice; &p , 0.05 versus IL-1RaDH mice, Kaplan–Meier statistic.

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Functional Assay, Derivative Assay, Control, Injection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Distinct roles of hepatocyte- and myeloid cell-derived IL-1 receptor antagonist during endotoxemia and sterile inflammation in mice.

doi: 10.4049/jimmunol.1000872

Figure Lengend Snippet: FIGURE 6. Contribution of hepatocytes and mye- loid cells as sources of circulating IL-1Ra after sys- temic challenge with IL-1b. IL-1Ra (A), IL-6 (B), CXCL1 (C), and CXCL2 (D) levels were quantified by ELISA in sera of control mice (WT), IL-1RaDH

Article Snippet: Endogenous peroxidase was blocked using 0.6% H2O2 for 10 min, and tissue sections were boiled in citrate-based Ag-unmasking solution (10 mM [pH 6]) for 3 min. To evaluate local expression of IL-1Ra, tissue sections were incubated with 14.4 mg/ml (for liver sections) or 7.2 mg/ml (for spleen sections) polyclonal goat anti-mouse IL-1Ra Ab (capture Ab of mouse IL-1Ra Duoset ELISA kit; R&D Systems) for 2 h. The sections were rinsed and incubated for 1 h with 1.6 mg/ml donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer Cell International

Article Title: MSCs loaded with oncolytic reovirus: migration and in vivo virus delivery potential for evaluating anti-cancer effect in tumor-bearing C57BL/6 mice

doi: 10.1186/s12935-021-01848-5

Figure Lengend Snippet: Specifications of all antibodies used in AD-MSCs immunophenotyping and cytokine assays

Article Snippet: Mouse TGF-β DuoSet ELISA , DY402-05 , R & D systems , Capture 4 μg/ml Detection 75 ng/ml.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer Cell International

Article Title: MSCs loaded with oncolytic reovirus: migration and in vivo virus delivery potential for evaluating anti-cancer effect in tumor-bearing C57BL/6 mice

doi: 10.1186/s12935-021-01848-5

Figure Lengend Snippet: Determination of the splenic IL-6, IFN-γ, IL-10, and TGF-β cytokine secretion in response to stimulators. According to analyzed data, the robust response in the released cytokines level was observed after 3rd treatments. Reovirus had a direct effect on released IFN-γ level and an indirect effect on inflammatory cytokines when accompanying with MSCs its function was moderated. The experiment was performed in triplicate (*: P ≤ 0.05; **: P ≤ 0.01)

Article Snippet: Mouse TGF-β DuoSet ELISA , DY402-05 , R & D systems , Capture 4 μg/ml Detection 75 ng/ml.

Techniques: