mouse anti human il 23p19  (R&D Systems)

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Specificity protein 1 (Sp1) production by human monocytes and macrophages after exposure to LPS Human monocytes and macrophages (on day 9 of culture) were stimulated with LPS (10 ng/mL) for 6 h Sp1 levels were measured by enzyme-linked immunosorbent assay (a). Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01 (with Bonferroni correction) Furthermore, the levels of Sp1were detected by Western blotting. The density of each band was normalized for that of GAPDH. A representative result is shown as arbitrary density units. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01, ∗ P < .05 (with Bonferroni correction) (b).

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of small interfering RNAs (siRNAs) for p22phox, gp91phox, dual oxidase 1 (DUOX1), and dual oxidase 2 (DUOX2) on specificity protein 1 (Sp1), IL-23, cyclooxygenase-2 (COX-2), and tumor necrosis factor α-induced protein 3 (TNFAIP3) after exposure to resiquimod Human macrophages (on day 9 of culture) transfected with siRNA for p22phox, gp91phox, DUOX1, and DUOX2 were stimulated with resiquimod (5μM) for 6 h, after which Sp1 and TNFAIP3 levels were measured by enzyme-linked immunosorbent assay (a and d). Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant Furthermore, levels of IL-23 (b) and COX-2 (c) were detected by Western blotting. The density of each band was normalized for that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A representative result is shown as arbitrary density units. Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant.

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of small interfering RNAs (siRNAs) for programmed cell death-1 (PDCD-1), hypoxia inducible factor 1α (HIF1α), triggering receptor expressed on myeloid cells 1 (TREM-1), and tumor necrosis factor α-induced protein 3 (TNFAIP3) on specificity protein 1 (Sp1) and IL-23 production after exposure to resiquimod Human macrophages (on day 9 of culture) transfected with small interfering RNA (siRNA) for PDCD-1, HIF1α, TREM-1, and TNFAIP3 were stimulated with resiquimod (5μM) for 6 h, after which Sp1 levels were measured by enzyme-linked immunosorbent assay. Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; (with Bonferroni correction); N.S., not significant.

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Transfection, Small Interfering RNA, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Tumor necrosis factor α-induced protein 3 (TNFAIP3) production by human monocytes and macrophages after exposure to lipopolysaccharide (LPS) or Pam3CsK4 Human monocytes and macrophages (on day 9 of culture) were stimulated with LPS (10 ng/mL) or Pam3CsK4 (1 μg/mL) for 6 h TNFAIP3 levels were measured by enzyme-linked immunosorbent assay. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction) (a). Furthermore, the levels of TNFAIP3 were detected by Western blotting. The density of each band was normalized for that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A representative result is shown. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01 (with Bonferroni correction); N.S., not significant (b).

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of a peroxisome proliferator-activated receptor α (PPARα) agonist (GW7647) and uncoupling protein 2 (UCP2) small interfering RNA (siRNA) on tumor necrosis factor α-induced protein 3 (TNFAIP3) and IL-23 production after exposure to resiquimod Human macrophages (on day 9 of culture) transfected or not with UCP2 siRNA were pretreated with a PPARα agonist (GW7647) for 1 h and then the cells were stimulated with resiquimod (5μM) for 6 h. Macrophages transfected with UCP2 siRNA and TNFAIP3 siRNA or pretreated with GW7647 for 1 h were stimulated with resiquimod (5μM) for 6 h, after which IL-23 levels were measured by enzyme-linked immunosorbent assay (a). Data were obtained by using samples from 3 individuals in each group; results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction) (a). Furthermore, TNFAIP3 levels were detected by Western blotting. The density of each band was normalized for that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A representative result is shown as arbitrary density units. Data were obtained by using samples from 3 individuals in each group; results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant (b).

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Small Interfering RNA, Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of small interfering RNAs (siRNAs) for p22phox, gp91phox, dual oxidase 1 (DUOX1), and dual oxidase 2 (DUOX2) on specificity protein 1 (Sp1), IL-23, cyclooxygenase-2 (COX-2), and tumor necrosis factor α-induced protein 3 (TNFAIP3) after exposure to resiquimod Human macrophages (on day 9 of culture) transfected with siRNA for p22phox, gp91phox, DUOX1, and DUOX2 were stimulated with resiquimod (5μM) for 6 h, after which Sp1 and TNFAIP3 levels were measured by enzyme-linked immunosorbent assay (a and d). Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant Furthermore, levels of IL-23 (b) and COX-2 (c) were detected by Western blotting. The density of each band was normalized for that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). A representative result is shown as arbitrary density units. Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant.

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of small interfering RNAs (siRNAs) for programmed cell death-1 (PDCD-1), hypoxia inducible factor 1α (HIF1α), triggering receptor expressed on myeloid cells 1 (TREM-1), and tumor necrosis factor α-induced protein 3 (TNFAIP3) on specificity protein 1 (Sp1) and IL-23 production after exposure to resiquimod Human macrophages (on day 9 of culture) transfected with small interfering RNA (siRNA) for PDCD-1, HIF1α, TREM-1, and TNFAIP3 were stimulated with resiquimod (5μM) for 6 h, after which Sp1 levels were measured by enzyme-linked immunosorbent assay. Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; (with Bonferroni correction); N.S., not significant.

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Transfection, Small Interfering RNA, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of TNFAIP3 and DUOX2 on IL-23 production after sequential exposure to lipopolysaccharide and resiquimod Human macrophages (on day 9 of culture) transfected with small interfering RNA (siRNA) for TNFAIP3 and DUOX2 were stimulated by sequential administration of lipopolysaccharide (LPS, 10 ng/mL) and resiquimod (5μM), after which IL-23 levels were measured by enzyme-linked immunosorbent assay. Data were obtained by using samples from 3 individuals in each group, and results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction).

Article Snippet: The membranes were incubated with 0.2 × 10 3 μg/L mouse anti-human IL-23p19, TNFAIP3, Sp1, TGFβ1, and COX-2 IgG (Santa Cruz Biotechnology) for 1 h at room temperature, washed and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Santa Cruz Biotechnology) diluted to 1:5000.

Techniques: Transfection, Small Interfering RNA, Enzyme-linked Immunosorbent Assay

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

Article Title: Aspergillus fumigatus-induced IL-22 is not restricted to a specific Th cell subset and is dependent on complement receptor 3.

doi: 10.4049/jimmunol.1202601

Figure Lengend Snippet: FIGURE 5. Role of IL-23 and IL-1 in A. fumigatus–induced IL-17A and IL-22. (A) IL-1b and IL-23 concentrations in PBMCs (2.5 3 106/ml) (n = 7) that were stimulated with HI A. fumigatus conidia or hyphae at 37C and 5% CO2 for 24 h. IL-17 and IL-22 were measured in the culture supernatants of PBMCs (2.5 3 106/ml) stimulated with HI A. fumigatus conidia in the presence or absence of 10 mg/ml IL-1Ra (B) (n = 7 for IL-17, n = 12 for IL-22), IL-23 (C) (n = 12), or anti–IL-23p19 (D) (n = 5 for IL-17, n = 6 for IL-22). Differences of the means were analyzed for significance using the Wilcoxon signed rank test. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: IL-1R signaling was blocked by its natural receptor antagonist (Ra) IL-1Ra (10 mg/ml) (Amgen, Thousand Oaks, CA), and IL-23 was blocked with mouse anti-human IL-23p19 (10 mg/ml) (R&D Systems).

Techniques: