mouse il 23  (Thermo Fisher)


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    Name:
    Mouse IL 23 Recombinant Protein
    Description:
    Mouse IL 23 Recombinant Protein for ELISA Ctrl FN
    Catalog Number:
    14-8231-63
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    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher mouse il 23
    CX 3 CR1 + MNP-derived TL1A synergizes with <t>IL-23</t> and IL-1β to induce IL-22 in intestinal ILC3. (A) Gene-set enrichment analysis was used to determine whether the indicated disease-related SNP were differentially expressed between CD14 + MNPs and CD103 + DCs. Significance was estimated using the hypergeometric cumulative distribution, with a raw p-value cutoff of 0.05 for differential expression. Data were averaged from two independent donors. (B) B cells (CD3 − CD19 + ), CX 3 CR1 + MNPs, CD103 + DCs, and Ly6C + monocytes were sorted from the intestinal lamina propria of Cx3cr1 GFP/+ and quantitative PCR for TL1A was performed. Relative quantitation was performed by ΔCt normalized to GAPDH expression. Data are from two biological replicates performed with two technical replicates. *, P ≤ 0.05. Two-tailed Student’s t test. (C–E) Sorted intestinal ILCs from mice (C and E) or cultured human intestinal ILCs (D) were stimulated with media alone, IL-1β, or IL-23 with or without TL1A as indicated for 18 h. Brefeldin was added to the cultures 4 h before intracellular cytokine staining for IL-22 (C and D) or GM-CSF (E). Data are representative of six independent experiments. (F–H) Sorted intestinal ILCs were transfected with siRNA targeting Tnfrsf25 or a scramble control. (F) Knockdown efficiency was assessed after 24 h by flow cytometry comparing scramble control (solid line) with Tnfrsf25 siRNA. One of two representative experiments is shown. ILCs were then cultured with media alone (-) or IL-23 and TL1A or co-cultured with CX 3 CR1 + MNPs with or without LPS as indicated for an additional 18 h. (G) IL-22 production was measured by intracellular flow cytometry. Brefeldin was added to the cultures 4 h before intracellular cytokine staining. Data are representative of two independent experiments. (H) IL-22 secretion by samples from G were assessed by ELISA, performed in duplicate, before addition of Brefeldin. *, P ≤ 0.05; **, P ≤ 0.01. Two-tailed Student’s t test. Error bars represent SEM.
    Mouse IL 23 Recombinant Protein for ELISA Ctrl FN
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    Images

    1) Product Images from "CX3CR1+ mononuclear phagocytes support colitis-associated innate lymphoid cell production of IL-22"

    Article Title: CX3CR1+ mononuclear phagocytes support colitis-associated innate lymphoid cell production of IL-22

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20140678

    CX 3 CR1 + MNP-derived TL1A synergizes with IL-23 and IL-1β to induce IL-22 in intestinal ILC3. (A) Gene-set enrichment analysis was used to determine whether the indicated disease-related SNP were differentially expressed between CD14 + MNPs and CD103 + DCs. Significance was estimated using the hypergeometric cumulative distribution, with a raw p-value cutoff of 0.05 for differential expression. Data were averaged from two independent donors. (B) B cells (CD3 − CD19 + ), CX 3 CR1 + MNPs, CD103 + DCs, and Ly6C + monocytes were sorted from the intestinal lamina propria of Cx3cr1 GFP/+ and quantitative PCR for TL1A was performed. Relative quantitation was performed by ΔCt normalized to GAPDH expression. Data are from two biological replicates performed with two technical replicates. *, P ≤ 0.05. Two-tailed Student’s t test. (C–E) Sorted intestinal ILCs from mice (C and E) or cultured human intestinal ILCs (D) were stimulated with media alone, IL-1β, or IL-23 with or without TL1A as indicated for 18 h. Brefeldin was added to the cultures 4 h before intracellular cytokine staining for IL-22 (C and D) or GM-CSF (E). Data are representative of six independent experiments. (F–H) Sorted intestinal ILCs were transfected with siRNA targeting Tnfrsf25 or a scramble control. (F) Knockdown efficiency was assessed after 24 h by flow cytometry comparing scramble control (solid line) with Tnfrsf25 siRNA. One of two representative experiments is shown. ILCs were then cultured with media alone (-) or IL-23 and TL1A or co-cultured with CX 3 CR1 + MNPs with or without LPS as indicated for an additional 18 h. (G) IL-22 production was measured by intracellular flow cytometry. Brefeldin was added to the cultures 4 h before intracellular cytokine staining. Data are representative of two independent experiments. (H) IL-22 secretion by samples from G were assessed by ELISA, performed in duplicate, before addition of Brefeldin. *, P ≤ 0.05; **, P ≤ 0.01. Two-tailed Student’s t test. Error bars represent SEM.
    Figure Legend Snippet: CX 3 CR1 + MNP-derived TL1A synergizes with IL-23 and IL-1β to induce IL-22 in intestinal ILC3. (A) Gene-set enrichment analysis was used to determine whether the indicated disease-related SNP were differentially expressed between CD14 + MNPs and CD103 + DCs. Significance was estimated using the hypergeometric cumulative distribution, with a raw p-value cutoff of 0.05 for differential expression. Data were averaged from two independent donors. (B) B cells (CD3 − CD19 + ), CX 3 CR1 + MNPs, CD103 + DCs, and Ly6C + monocytes were sorted from the intestinal lamina propria of Cx3cr1 GFP/+ and quantitative PCR for TL1A was performed. Relative quantitation was performed by ΔCt normalized to GAPDH expression. Data are from two biological replicates performed with two technical replicates. *, P ≤ 0.05. Two-tailed Student’s t test. (C–E) Sorted intestinal ILCs from mice (C and E) or cultured human intestinal ILCs (D) were stimulated with media alone, IL-1β, or IL-23 with or without TL1A as indicated for 18 h. Brefeldin was added to the cultures 4 h before intracellular cytokine staining for IL-22 (C and D) or GM-CSF (E). Data are representative of six independent experiments. (F–H) Sorted intestinal ILCs were transfected with siRNA targeting Tnfrsf25 or a scramble control. (F) Knockdown efficiency was assessed after 24 h by flow cytometry comparing scramble control (solid line) with Tnfrsf25 siRNA. One of two representative experiments is shown. ILCs were then cultured with media alone (-) or IL-23 and TL1A or co-cultured with CX 3 CR1 + MNPs with or without LPS as indicated for an additional 18 h. (G) IL-22 production was measured by intracellular flow cytometry. Brefeldin was added to the cultures 4 h before intracellular cytokine staining. Data are representative of two independent experiments. (H) IL-22 secretion by samples from G were assessed by ELISA, performed in duplicate, before addition of Brefeldin. *, P ≤ 0.05; **, P ≤ 0.01. Two-tailed Student’s t test. Error bars represent SEM.

    Techniques Used: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Quantitation Assay, Two Tailed Test, Mouse Assay, Cell Culture, Staining, Transfection, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Increased ILC3 production of IL-22 in mild to moderate IBD correlates with presence of fecal stream. (A) LPMCs isolated from descending colon biopsies from patients with endoscopically mild to moderate Crohn’s’ disease ( n = 8, gray) or ulcerative colitis ( n = 6, black; Table S1 ), as well as age-matched non-IBD control patients undergoing routine screening colonoscopy ( n = 8, white), were stimulated ex vivo with PMA/ionomycin and evaluated by intracellular cytokine staining for expression of IL-17 and IL-22. The percentage of CD3 + or CD3 − fraction expressing IL-17 or IL-22 is indicated. *, P ≤ 0.05, two-tailed Student’s t test. Black bars represent the geometric mean. (B) Expression of c-Kit and CD56 in electronically gated CD3 − IL-22 + (black lines) and CD3 − IL-22 − (gray) LPMCs. (C) Expression of RORγt by c-Kit + CD56 + LPMCs. Lin − cells (CD14/CD19/CD3/CD11b/CD11c/TCRγδ − ; Fig. S3 A ) were stained with antibodies to surface markers c-Kit and CD56 and to intracellular RORγt. Lin − CD56 + c-Kit + ILC3 (black line) were compared with Lin − CD56 + c-Kit − NK cells (gray) for RORγt expression. (D) Surface staining of Lin − c-Kit + ILCs for the indicated markers (black line) compared with isotype control (gray) and all live LPMCs (dotted line) (E) LPMCs stained for intracellular IL-22 after stimulation with IL-23 for 3 h (solid line) or with control media (dotted line). Cells shown were gated on Lin − CD56 + c-Kit + . The isotype control is in gray. (F) CD11c + MHCII + human colonic APCs were electronically gated for expression of CD103 and CD14. One of three donors is shown. (G) Lamina propria cells from biopsy samples of tissue exposed (prediversion) or not exposed to the fecal stream (post-diversion) were cultured for 3 h and ILC3 production of IL-22 was assessed by flow cytometry. (left) Result from one representative donor. (right) Percentage of IL-22 + ILCs in afferent (Pre) and efferent (Post) limbs of three diverted patients. **, P ≤ 0.01, two-tailed Student’s t test. Black bars represent the geometric mean.
    Figure Legend Snippet: Increased ILC3 production of IL-22 in mild to moderate IBD correlates with presence of fecal stream. (A) LPMCs isolated from descending colon biopsies from patients with endoscopically mild to moderate Crohn’s’ disease ( n = 8, gray) or ulcerative colitis ( n = 6, black; Table S1 ), as well as age-matched non-IBD control patients undergoing routine screening colonoscopy ( n = 8, white), were stimulated ex vivo with PMA/ionomycin and evaluated by intracellular cytokine staining for expression of IL-17 and IL-22. The percentage of CD3 + or CD3 − fraction expressing IL-17 or IL-22 is indicated. *, P ≤ 0.05, two-tailed Student’s t test. Black bars represent the geometric mean. (B) Expression of c-Kit and CD56 in electronically gated CD3 − IL-22 + (black lines) and CD3 − IL-22 − (gray) LPMCs. (C) Expression of RORγt by c-Kit + CD56 + LPMCs. Lin − cells (CD14/CD19/CD3/CD11b/CD11c/TCRγδ − ; Fig. S3 A ) were stained with antibodies to surface markers c-Kit and CD56 and to intracellular RORγt. Lin − CD56 + c-Kit + ILC3 (black line) were compared with Lin − CD56 + c-Kit − NK cells (gray) for RORγt expression. (D) Surface staining of Lin − c-Kit + ILCs for the indicated markers (black line) compared with isotype control (gray) and all live LPMCs (dotted line) (E) LPMCs stained for intracellular IL-22 after stimulation with IL-23 for 3 h (solid line) or with control media (dotted line). Cells shown were gated on Lin − CD56 + c-Kit + . The isotype control is in gray. (F) CD11c + MHCII + human colonic APCs were electronically gated for expression of CD103 and CD14. One of three donors is shown. (G) Lamina propria cells from biopsy samples of tissue exposed (prediversion) or not exposed to the fecal stream (post-diversion) were cultured for 3 h and ILC3 production of IL-22 was assessed by flow cytometry. (left) Result from one representative donor. (right) Percentage of IL-22 + ILCs in afferent (Pre) and efferent (Post) limbs of three diverted patients. **, P ≤ 0.01, two-tailed Student’s t test. Black bars represent the geometric mean.

    Techniques Used: Isolation, Ex Vivo, Staining, Expressing, Two Tailed Test, Cell Culture, Flow Cytometry, Cytometry

    TLR-stimulated CX 3 CR1 + MNPs are stronger inducers of ILC3 production of IL-22 than CD103 + CD11b + DCs and monocytes. (A–C) CD103 or CX 3 CR1 + MHCII + CD11c + CD11b + cells were isolated from the lamina propria of CX 3 CR1 GFP/+ mice (sort strategy shown in A and co-cultured with Lin − RORγt-GFP + ILCs with or without the indicated bacterial TLR ligands or IL-23. IL-22 was assessed by intracellular staining of CD90.2 + ILCs after 18 h. A representative flow cytometry plot is shown in B. (C) Percent IL-22 + ILCs is shown from seven independent experiments. **, P ≤ 0.01; *, P ≤ 0.05. One way ANOVA with Bonferroni’s correction. Error bars represent SEM. (D–F) Ly6C + MHCII lo (monocytes) and Ly6C − MHCII hi (MNPs) were isolated from CX 3 CR1 + CD11b + lamina propria cells (sort strategy is shown in D and co-cultured with intestinal ILCs with LPS or IL-23 as indicated. Intracellular cytokine staining for IL-22 is shown after 18 h (E). Supernatants were harvested after 18 h and IL-22 production quantitated by ELISA (F). Results are representative of two independent experiments performed in triplicate. ***, P ≤ 0.001. One-way ANOVA with Bonferroni correction. Error bars represent the SEM.
    Figure Legend Snippet: TLR-stimulated CX 3 CR1 + MNPs are stronger inducers of ILC3 production of IL-22 than CD103 + CD11b + DCs and monocytes. (A–C) CD103 or CX 3 CR1 + MHCII + CD11c + CD11b + cells were isolated from the lamina propria of CX 3 CR1 GFP/+ mice (sort strategy shown in A and co-cultured with Lin − RORγt-GFP + ILCs with or without the indicated bacterial TLR ligands or IL-23. IL-22 was assessed by intracellular staining of CD90.2 + ILCs after 18 h. A representative flow cytometry plot is shown in B. (C) Percent IL-22 + ILCs is shown from seven independent experiments. **, P ≤ 0.01; *, P ≤ 0.05. One way ANOVA with Bonferroni’s correction. Error bars represent SEM. (D–F) Ly6C + MHCII lo (monocytes) and Ly6C − MHCII hi (MNPs) were isolated from CX 3 CR1 + CD11b + lamina propria cells (sort strategy is shown in D and co-cultured with intestinal ILCs with LPS or IL-23 as indicated. Intracellular cytokine staining for IL-22 is shown after 18 h (E). Supernatants were harvested after 18 h and IL-22 production quantitated by ELISA (F). Results are representative of two independent experiments performed in triplicate. ***, P ≤ 0.001. One-way ANOVA with Bonferroni correction. Error bars represent the SEM.

    Techniques Used: Isolation, Mouse Assay, Cell Culture, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    CX 3 CR1 + MNP-derived IL-23 and IL-1β activate ILC3 to produce IL-22. (A) Phenotype analysis of colonic LPMCs from Cx3cr1 STOP-DTR/GFP mice with or without CD11c-cre after DT injection for 2 d. (top) Selective depletion of CX 3 CR1 hi MNPs. (bottom) Expression of Ly6C and MHCII on CX 3 CR1 hi and CX 3 CR1 int populations. (B) Expression of IL-22 in Lin − CD90.2 + colonic ILCs from Cx3cr1 STOP-DTR/+ (Stop-DTR) or CD11c-Cre x Cx3cr1 STOP-DTR/+ (Cre-DTR) mice at 7 d after C. rodentium infection. DT was administered at days −2, −1, and 0 and every other day postinfection. One representative intracellular cytokine flow cytometry plot is shown on the left and a composite graph ( n = 6/group) on the right. *, P ≤ 0.05, two-tailed Student’s t test. Error bars represent the SEM. Results are a composite of two independent experiments. (C) Supernatants from APC-ILC co-cultures ( Fig. 3, B and C ) were harvested after 18 h and assayed for IL-23 by ELISA. Results are averages of three independent experiments and the SEM is shown. (D) CX 3 CR1 + MNPs or CD103 + CD11b + DCs were sorted and incubated with media or CpG for 18 h and supernatants were assayed for IL-1β by ELISA. Results are the mean of two independent experiments performed in duplicate and the SEM is shown. *, P ≤ 0.05; **, P ≤ 0.01. (E) Lin − CD90.2 hi ILCs from WT or Il1r −/− mice were co-cultured with sorted intestinal MNPs from WT or Il23p19 −/− mice, with or without CpG, as indicated. IL-22 production by the ILCs was assessed after 18 h by ELISA. Data are combined from three independent experiments performed in duplicate. *, P ≤ 0.05; ***, P ≤ 0.001. One-way ANOVA with Bonferroni correction. Error bars represent the SEM.
    Figure Legend Snippet: CX 3 CR1 + MNP-derived IL-23 and IL-1β activate ILC3 to produce IL-22. (A) Phenotype analysis of colonic LPMCs from Cx3cr1 STOP-DTR/GFP mice with or without CD11c-cre after DT injection for 2 d. (top) Selective depletion of CX 3 CR1 hi MNPs. (bottom) Expression of Ly6C and MHCII on CX 3 CR1 hi and CX 3 CR1 int populations. (B) Expression of IL-22 in Lin − CD90.2 + colonic ILCs from Cx3cr1 STOP-DTR/+ (Stop-DTR) or CD11c-Cre x Cx3cr1 STOP-DTR/+ (Cre-DTR) mice at 7 d after C. rodentium infection. DT was administered at days −2, −1, and 0 and every other day postinfection. One representative intracellular cytokine flow cytometry plot is shown on the left and a composite graph ( n = 6/group) on the right. *, P ≤ 0.05, two-tailed Student’s t test. Error bars represent the SEM. Results are a composite of two independent experiments. (C) Supernatants from APC-ILC co-cultures ( Fig. 3, B and C ) were harvested after 18 h and assayed for IL-23 by ELISA. Results are averages of three independent experiments and the SEM is shown. (D) CX 3 CR1 + MNPs or CD103 + CD11b + DCs were sorted and incubated with media or CpG for 18 h and supernatants were assayed for IL-1β by ELISA. Results are the mean of two independent experiments performed in duplicate and the SEM is shown. *, P ≤ 0.05; **, P ≤ 0.01. (E) Lin − CD90.2 hi ILCs from WT or Il1r −/− mice were co-cultured with sorted intestinal MNPs from WT or Il23p19 −/− mice, with or without CpG, as indicated. IL-22 production by the ILCs was assessed after 18 h by ELISA. Data are combined from three independent experiments performed in duplicate. *, P ≤ 0.05; ***, P ≤ 0.001. One-way ANOVA with Bonferroni correction. Error bars represent the SEM.

    Techniques Used: Derivative Assay, Mouse Assay, Injection, Expressing, Infection, Flow Cytometry, Cytometry, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Incubation, Cell Culture

    Human ILC3 production of IL-22 is supported by IL-23 and IL-1β produced by TLR-stimulated CD14 + and CX 3 CR1 + MNPs. (A–C) HLA-DR + CD11c + cells from intestinal resection tissue were sorted into CD103 + DCs and CD14 + MNPs subpopulations and transcriptional profiles were assessed by RNA-seq. (A) Sorting strategy. (B) Each subset was examined for expression of the indicated cell surface markers. Isotype controls are shown in gray. One of three donors is shown. (C) Heatmap of relative expression of relevant MNP-related genes. Values represent the mean of two independent donors, and an asterisk denotes individual genes differentially expressed at an FDR = 0.01. (D and E) Induction of IL-22 in human ILCs in co-culture with CD14 + MNPs or CD103 + DCs in the presence of media alone, LPS, or flagellin, as indicated. c-Kit + cells were examined for intracellular IL-22 production after 18-h culture. Data are representative of five independent experiments. (F) CD14 + MNPs or CD103 + DCs sorted from human intestine (as in A) were stimulated with the indicated TLR ligands for 18 h and qPCR or cytometric bead array analysis were used to quantitate IL-23p19 and IL-1β, respectively. Results are averaged from three independent donors, and technical replicates were performed in duplicate or triplicate, respectively. *, P ≤ 0.05. N.D., not detected. Two-tailed Student’s t test. Error bars represent the SEM. (G) Sorted human intestinal CD14 + MNPs and ILCs were left unstimulated or were co-cultured in the presence of LPS with or without neutralizing antibodies against IL-1β and IL-23. IL-22 ELISA was performed after 18h. Results are averaged from two independent donors performed in duplicate. *, P ≤ 0.05. Two-tailed Student’s t test. Error bars represent the SEM.
    Figure Legend Snippet: Human ILC3 production of IL-22 is supported by IL-23 and IL-1β produced by TLR-stimulated CD14 + and CX 3 CR1 + MNPs. (A–C) HLA-DR + CD11c + cells from intestinal resection tissue were sorted into CD103 + DCs and CD14 + MNPs subpopulations and transcriptional profiles were assessed by RNA-seq. (A) Sorting strategy. (B) Each subset was examined for expression of the indicated cell surface markers. Isotype controls are shown in gray. One of three donors is shown. (C) Heatmap of relative expression of relevant MNP-related genes. Values represent the mean of two independent donors, and an asterisk denotes individual genes differentially expressed at an FDR = 0.01. (D and E) Induction of IL-22 in human ILCs in co-culture with CD14 + MNPs or CD103 + DCs in the presence of media alone, LPS, or flagellin, as indicated. c-Kit + cells were examined for intracellular IL-22 production after 18-h culture. Data are representative of five independent experiments. (F) CD14 + MNPs or CD103 + DCs sorted from human intestine (as in A) were stimulated with the indicated TLR ligands for 18 h and qPCR or cytometric bead array analysis were used to quantitate IL-23p19 and IL-1β, respectively. Results are averaged from three independent donors, and technical replicates were performed in duplicate or triplicate, respectively. *, P ≤ 0.05. N.D., not detected. Two-tailed Student’s t test. Error bars represent the SEM. (G) Sorted human intestinal CD14 + MNPs and ILCs were left unstimulated or were co-cultured in the presence of LPS with or without neutralizing antibodies against IL-1β and IL-23. IL-22 ELISA was performed after 18h. Results are averaged from two independent donors performed in duplicate. *, P ≤ 0.05. Two-tailed Student’s t test. Error bars represent the SEM.

    Techniques Used: Produced, RNA Sequencing Assay, Expressing, Co-Culture Assay, Real-time Polymerase Chain Reaction, Two Tailed Test, Cell Culture, Enzyme-linked Immunosorbent Assay

    2) Product Images from "CCR6 is required for IL-23-induced psoriasis-like inflammation in mice"

    Article Title: CCR6 is required for IL-23-induced psoriasis-like inflammation in mice

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI37378

    IL-23–injected ears of WT and Ccr6 –/– mice have similar IL17a and Il17f mRNA, Th17 cells, and IL-22–producing cells at day 15.
    Figure Legend Snippet: IL-23–injected ears of WT and Ccr6 –/– mice have similar IL17a and Il17f mRNA, Th17 cells, and IL-22–producing cells at day 15.

    Techniques Used: Injection, Mouse Assay

    Il22 mRNA correlates with IL-23–induced inflammation, but not with Il17a and Il17f mRNA, and is produced in T cell–independent and T cell–dependent phases.
    Figure Legend Snippet: Il22 mRNA correlates with IL-23–induced inflammation, but not with Il17a and Il17f mRNA, and is produced in T cell–independent and T cell–dependent phases.

    Techniques Used: Produced

    IL-23 injections increase the number of CD4 + T cells and dendritic cells in ears of WT mice, but not Ccr6 –/– mice.
    Figure Legend Snippet: IL-23 injections increase the number of CD4 + T cells and dendritic cells in ears of WT mice, but not Ccr6 –/– mice.

    Techniques Used: Mouse Assay

    Rag1 –/– and WT mice show similar initial responses to injections with IL-23.
    Figure Legend Snippet: Rag1 –/– and WT mice show similar initial responses to injections with IL-23.

    Techniques Used: Mouse Assay

    Ccr6 –/– mice are resistant to IL-23–induced acanthosis and dermal inflammation.
    Figure Legend Snippet: Ccr6 –/– mice are resistant to IL-23–induced acanthosis and dermal inflammation.

    Techniques Used: Mouse Assay

    3) Product Images from "CCR6 is required for IL-23-induced psoriasis-like inflammation in mice"

    Article Title: CCR6 is required for IL-23-induced psoriasis-like inflammation in mice

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI37378

    IL-23–injected ears of WT and Ccr6 –/– mice have similar IL17a and Il17f mRNA, Th17 cells, and IL-22–producing cells at day 15.
    Figure Legend Snippet: IL-23–injected ears of WT and Ccr6 –/– mice have similar IL17a and Il17f mRNA, Th17 cells, and IL-22–producing cells at day 15.

    Techniques Used: Injection, Mouse Assay

    Il22 mRNA correlates with IL-23–induced inflammation, but not with Il17a and Il17f mRNA, and is produced in T cell–independent and T cell–dependent phases.
    Figure Legend Snippet: Il22 mRNA correlates with IL-23–induced inflammation, but not with Il17a and Il17f mRNA, and is produced in T cell–independent and T cell–dependent phases.

    Techniques Used: Produced

    IL-23 injections increase the number of CD4 + T cells and dendritic cells in ears of WT mice, but not Ccr6 –/– mice.
    Figure Legend Snippet: IL-23 injections increase the number of CD4 + T cells and dendritic cells in ears of WT mice, but not Ccr6 –/– mice.

    Techniques Used: Mouse Assay

    Rag1 –/– and WT mice show similar initial responses to injections with IL-23.
    Figure Legend Snippet: Rag1 –/– and WT mice show similar initial responses to injections with IL-23.

    Techniques Used: Mouse Assay

    Ccr6 –/– mice are resistant to IL-23–induced acanthosis and dermal inflammation.
    Figure Legend Snippet: Ccr6 –/– mice are resistant to IL-23–induced acanthosis and dermal inflammation.

    Techniques Used: Mouse Assay

    4) Product Images from "Prostaglandin E2-dependent IL-23 production in aged murine dendritic cells"

    Article Title: Prostaglandin E2-dependent IL-23 production in aged murine dendritic cells

    Journal: Experimental gerontology

    doi: 10.1016/j.exger.2010.06.007

    Expression of Th17-favorable mediators (Compounds) IL-23, PGE2 and TGF-β is significantly increased in aged DCs upon TLR activation
    Figure Legend Snippet: Expression of Th17-favorable mediators (Compounds) IL-23, PGE2 and TGF-β is significantly increased in aged DCs upon TLR activation

    Techniques Used: Expressing, Activation Assay

    PGE2 upregulates IL-23 production from aged DCs
    Figure Legend Snippet: PGE2 upregulates IL-23 production from aged DCs

    Techniques Used:

    5) Product Images from "Increased Th17 cells in the tumor microenvironment is mediated by IL-23 via tumor-secreted PGE2"

    Article Title: Increased Th17 cells in the tumor microenvironment is mediated by IL-23 via tumor-secreted PGE2

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

    doi: 10.4049/jimmunol.1203141

    Tumor conditioned medium increases IL-23 production in DCs and macrophages. 1 × 10 6 Flt3 ligand-induced immature DCs were purified and seeded in each well of a 24-well culture plate. Different amounts of 4T1 TCM as indicated were added to the
    Figure Legend Snippet: Tumor conditioned medium increases IL-23 production in DCs and macrophages. 1 × 10 6 Flt3 ligand-induced immature DCs were purified and seeded in each well of a 24-well culture plate. Different amounts of 4T1 TCM as indicated were added to the

    Techniques Used: Purification

    IL-23 expression is increased in breast cancer. Total RNA was extracted from tumor-surrounding tissues and breast cancer tissues from human patients ( A–C ) and tumor-bearing mice ( D–F ). Quantitative mRNA expression of human IL-23 p19 (
    Figure Legend Snippet: IL-23 expression is increased in breast cancer. Total RNA was extracted from tumor-surrounding tissues and breast cancer tissues from human patients ( A–C ) and tumor-bearing mice ( D–F ). Quantitative mRNA expression of human IL-23 p19 (

    Techniques Used: Expressing, Mouse Assay

    Tumor-derived PGE 2 enhances IL-23 expression at the transcriptional level through binding to the CRE site in the p19 promoter. 3 × 10 6 RAW cells were stimulated with LPS (1 μg/ml) or LPS plus 400 μl 4T1 TCM ( A ), or LPS plus PGE
    Figure Legend Snippet: Tumor-derived PGE 2 enhances IL-23 expression at the transcriptional level through binding to the CRE site in the p19 promoter. 3 × 10 6 RAW cells were stimulated with LPS (1 μg/ml) or LPS plus 400 μl 4T1 TCM ( A ), or LPS plus PGE

    Techniques Used: Derivative Assay, Expressing, Binding Assay

    Tumor-secreted PGE 2 enhances IL-23 production through EP2 and EP4 receptors. 1 × 10 6 RAW cells were cultured with normal media (NM), 400 μl/ml of 4T1 TCM, or 400 μl/ml of TCM collected from 4T1 cells treated with NS398 in the presence
    Figure Legend Snippet: Tumor-secreted PGE 2 enhances IL-23 production through EP2 and EP4 receptors. 1 × 10 6 RAW cells were cultured with normal media (NM), 400 μl/ml of 4T1 TCM, or 400 μl/ml of TCM collected from 4T1 cells treated with NS398 in the presence

    Techniques Used: Cell Culture

    6) Product Images from "Withasteroid B from D. metel L. regulates immune responses by modulating the JAK/STAT pathway and the IL‐17+RORγt+/IL‐10+FoxP3+ ratio"

    Article Title: Withasteroid B from D. metel L. regulates immune responses by modulating the JAK/STAT pathway and the IL‐17+RORγt+/IL‐10+FoxP3+ ratio

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/cei.12998

    B2 ameliorates cutaneous inflammation in a mouse model. (a,b) Mouse ears were treated topically with B2 and subsequently injected intradermally with phosphate‐buffered saline (PBS) as a vehicle or mouse interleukin (IL)‐23 every other day for 14 days ( n = 6 mice/group). Ear swelling was evaluated 24 h after the final injection. Histology of mouse ear sections was assessed by haematoxylin and eosin (H E) staining. Acanthosis (c) and dermal cellular infiltrates (d) of skin injected with PBS or B2 collected from mice injected with IL‐23. * P
    Figure Legend Snippet: B2 ameliorates cutaneous inflammation in a mouse model. (a,b) Mouse ears were treated topically with B2 and subsequently injected intradermally with phosphate‐buffered saline (PBS) as a vehicle or mouse interleukin (IL)‐23 every other day for 14 days ( n = 6 mice/group). Ear swelling was evaluated 24 h after the final injection. Histology of mouse ear sections was assessed by haematoxylin and eosin (H E) staining. Acanthosis (c) and dermal cellular infiltrates (d) of skin injected with PBS or B2 collected from mice injected with IL‐23. * P

    Techniques Used: Injection, Mouse Assay, Staining

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    Expressing:

    Article Title: Identification of Canonical Tyrosine-dependent and Non-canonical Tyrosine-independent STAT3 Activation Sites in the Intracellular Domain of the Interleukin 23 Receptor *
    Article Snippet: .. After selection and proliferation analysis, Ba/F3-gp130 cells expressing murine IL-23R variants and murine IL-12Rβ1 were cultured in the respective medium containing 10 ng/ml recombinant mouse IL-23 as described (eBioscience, San Diego, CA) or Hyper-IL-23 (HIL-23) instead of HIL-6. ..

    Cell Culture:

    Article Title: Identification of Canonical Tyrosine-dependent and Non-canonical Tyrosine-independent STAT3 Activation Sites in the Intracellular Domain of the Interleukin 23 Receptor *
    Article Snippet: .. After selection and proliferation analysis, Ba/F3-gp130 cells expressing murine IL-23R variants and murine IL-12Rβ1 were cultured in the respective medium containing 10 ng/ml recombinant mouse IL-23 as described (eBioscience, San Diego, CA) or Hyper-IL-23 (HIL-23) instead of HIL-6. ..

    Recombinant:

    Article Title: Identification of Canonical Tyrosine-dependent and Non-canonical Tyrosine-independent STAT3 Activation Sites in the Intracellular Domain of the Interleukin 23 Receptor *
    Article Snippet: .. After selection and proliferation analysis, Ba/F3-gp130 cells expressing murine IL-23R variants and murine IL-12Rβ1 were cultured in the respective medium containing 10 ng/ml recombinant mouse IL-23 as described (eBioscience, San Diego, CA) or Hyper-IL-23 (HIL-23) instead of HIL-6. ..

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?
    Article Snippet: Cytokine secretion by splenocytes or pancreatic draining lymph node cells was determined by ELISA as recommended by the kit manufacturer (eBioscience). .. Briefly, cells from female NOD mice (5 × 105 ) were incubated in 96-well flat-bottomed microtitre plates with 0·1 μg/ml of anti-CD3 antibody (1452C11) in the presence or absence of 10 ng/ml of recombinant mouse IL-23 (eBioscience) and the supernatants were harvested after 48 hr. .. Levels of IL-17, IFN-γ, TGF-β and IL-10 were determined in triplicate in 0·1 ml of supernatant using a sandwich ELISA method.

    Article Title: Macrophage Migration Inhibitory Factor (MIF) Drives Murine Psoriasiform Dermatitis
    Article Snippet: Induction of recombinant IL-23-induced dermatitis IL-23-induced dermatitis was induced and evaluated, as previously described ( ). .. Briefly, 0.5 μg of recombinant murine IL-23 (eBioscience, Frankfurt, Germany), solved in 1% BSA in PBS, was injected i.d. in a volume of 20 μl into the dorsal surface of right ear, vehicle control (1% BSA in PBS) into the left ear of recipient mice every other day, starting day 0 of the experiment. .. Generation of bone marrow chimera Bone marrow chimera were generated, as described previously ( ).

    Article Title: Microbiota-induced TNF-like ligand 1A drives group 3 innate lymphoid cell-mediated barrier protection and intestinal T cell activation during colitis
    Article Snippet: Lineage- KLRG1- IL23R-GFP mouse ILC3 were sorted from LPMCs and resuspended in RPMI (10% FBS) tissue culture media for stimulation directly ex vivo. .. Mouse ILCs were stimulated with mouse recombinant IL-23 (eBioscience; 40 ng/ml), IL-1 (eBioscience; 10 ng/ml), or TL1A (R & D Systems; 200 ng/ml), as indicated. .. After 18 h, supernatants were harvested for IL-22 or GM-CSF ELISA (eBioscience) and Golgi Plug (BD) was added to cells for 4 h for subsequent intracellular cytokine staining.

    Article Title: Early MyD88-Dependent Induction of Interleukin-17A Expression during Salmonella Colitis ▿ Colitis ▿ †
    Article Snippet: Cells were washed and placed in complete IMDM containing 25 μg/ml of gentamicin for an additional 3 h. Isolation of untouched splenic T cells was performed using a pan-T-cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) per the manufacturer's instructions. .. Cells were treated with 10 ng/ml of recombinant mouse IL-23 and/or 10 ng/ml of IL-1β (eBioscience, San Diego, CA), for a total of 5 h. RNA was isolated using a Qiagen Qiashredder/RNEasy kit per the manufacturer's instructions. .. Three groups of two naive mice each (C57BL/6; Taconic) were euthanized, and organ tissue samples from each group of mice were combined.

    Article Title: Anti-inflammatory activity of compounds isolated from Astragalus sinicus L. in cytokine-induced keratinocytes and skin
    Article Snippet: Recombinant mouse cytokines against interleukin (IL)-2, IL-4, IL-6 and IL-12, and mouse antibodies against anti-IL-4, anti-IFN-γ and anti-CD28 were purchased from BD Biosciences (San Jose, CA, USA). .. Recombinant human TNF-α, IFN-γ and TGF-β were obtained from PROSPEC (East Brunswick, NJ, USA) and recombinant mouse IL-23 was obtained from eBioscience (San Diego, CA, USA). .. Antibodies specific for phospho-IκBα (Ser32), phospho-NF-κB p65 (Ser536), phospho-STAT1 (Tyr701), STAT1, phospho-STAT3 (Tyr705), phospho-Akt (Ser473), Akt, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-p38 (Thr180/Tyr182), p38, phospho-JNK (Thr183/Tyr185), JNK, phospho-Src (Tyr416), Src, phospho-Lck (Tyr505), Lck, phospho-Lyn (Tyr507), Lyn, cyclooxygenase-2 (COX-2), intercellular adhesion molecule 1 (ICAM-1), PARP, cleaved caspase-3 (Asp175), cleaved caspase-9 (Asp330), p21, p27, Bax and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA), and antibodies specific for IκBα, NF-κB p65, STAT3 and Lamin B1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Mouse Assay:

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?
    Article Snippet: Cytokine secretion by splenocytes or pancreatic draining lymph node cells was determined by ELISA as recommended by the kit manufacturer (eBioscience). .. Briefly, cells from female NOD mice (5 × 105 ) were incubated in 96-well flat-bottomed microtitre plates with 0·1 μg/ml of anti-CD3 antibody (1452C11) in the presence or absence of 10 ng/ml of recombinant mouse IL-23 (eBioscience) and the supernatants were harvested after 48 hr. .. Levels of IL-17, IFN-γ, TGF-β and IL-10 were determined in triplicate in 0·1 ml of supernatant using a sandwich ELISA method.

    Article Title: Macrophage Migration Inhibitory Factor (MIF) Drives Murine Psoriasiform Dermatitis
    Article Snippet: Induction of recombinant IL-23-induced dermatitis IL-23-induced dermatitis was induced and evaluated, as previously described ( ). .. Briefly, 0.5 μg of recombinant murine IL-23 (eBioscience, Frankfurt, Germany), solved in 1% BSA in PBS, was injected i.d. in a volume of 20 μl into the dorsal surface of right ear, vehicle control (1% BSA in PBS) into the left ear of recipient mice every other day, starting day 0 of the experiment. .. Generation of bone marrow chimera Bone marrow chimera were generated, as described previously ( ).

    Incubation:

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?
    Article Snippet: Cytokine secretion by splenocytes or pancreatic draining lymph node cells was determined by ELISA as recommended by the kit manufacturer (eBioscience). .. Briefly, cells from female NOD mice (5 × 105 ) were incubated in 96-well flat-bottomed microtitre plates with 0·1 μg/ml of anti-CD3 antibody (1452C11) in the presence or absence of 10 ng/ml of recombinant mouse IL-23 (eBioscience) and the supernatants were harvested after 48 hr. .. Levels of IL-17, IFN-γ, TGF-β and IL-10 were determined in triplicate in 0·1 ml of supernatant using a sandwich ELISA method.

    Injection:

    Article Title: Macrophage Migration Inhibitory Factor (MIF) Drives Murine Psoriasiform Dermatitis
    Article Snippet: Induction of recombinant IL-23-induced dermatitis IL-23-induced dermatitis was induced and evaluated, as previously described ( ). .. Briefly, 0.5 μg of recombinant murine IL-23 (eBioscience, Frankfurt, Germany), solved in 1% BSA in PBS, was injected i.d. in a volume of 20 μl into the dorsal surface of right ear, vehicle control (1% BSA in PBS) into the left ear of recipient mice every other day, starting day 0 of the experiment. .. Generation of bone marrow chimera Bone marrow chimera were generated, as described previously ( ).

    Isolation:

    Article Title: Early MyD88-Dependent Induction of Interleukin-17A Expression during Salmonella Colitis ▿ Colitis ▿ †
    Article Snippet: Cells were washed and placed in complete IMDM containing 25 μg/ml of gentamicin for an additional 3 h. Isolation of untouched splenic T cells was performed using a pan-T-cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) per the manufacturer's instructions. .. Cells were treated with 10 ng/ml of recombinant mouse IL-23 and/or 10 ng/ml of IL-1β (eBioscience, San Diego, CA), for a total of 5 h. RNA was isolated using a Qiagen Qiashredder/RNEasy kit per the manufacturer's instructions. .. Three groups of two naive mice each (C57BL/6; Taconic) were euthanized, and organ tissue samples from each group of mice were combined.

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  • 93
    Thermo Fisher recombinant il23
    Targeted disruption of SOCS3 promotes activation of BMP2-Smad signaling. (A) MC3T3/E1 cells were treated with BMP2 (100 ng/mL), and the phosphorylation level of Smad protein was examined by Western blotting. Shown are representative blots from three independent experiments. (B) SOCS3 knockdown and control cell lines were generated by lentivirus infection of MC3T3/E1 cells. The cells were then lysed and SOCS3 knockdown efficiency was examined by Western blotting. Shown are representative blots from three independent experiments. (C) SOCS3 knockdown and control cell lines were treated with BMP2 (100 ng/mL) for 0, 5, 15, 30 min and the phosphorylation level of Smad protein was detected by Western blotting. Shown are representative blots from three independent experiments. (D) BMSCs harvested from SOCS3 KD and WT mice were treated with BMP2 (100 ng/mL) or / and <t>IL23</t> (100 ng/mL) cytokines. The cells were stimulated for 21 days and then stained with alizarin red S. Shown are representative photomicrographs.
    Recombinant Il23, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tnf α
    Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, <t>TNF-α,</t> IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p
    Tnf α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher il 23
    Role played by interleukin <t>(IL)–23.</t> Groups of mice were intranasally inoculated with 2 × 10 6 yeast cells, and lungs were analyzed at serial intervals for expression of IL-23 by quantitative reverse-transcription polymerase chain reaction
    Il 23, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher interleukin 2
    Analysis of T-cell activation after immunization with the RNA adjuvant and MERS S protein vaccine. ( A ) The population of MERS S protein-specific cells secreting interferon-γ (IFN-γ) and <t>interleukin-2</t> (IL-2) were quantified by using ELISPOT assay, after treatment with MERS S protein in cultured splenocytes from immunized mice. ( B ) Cytokine levels in splenocyte supernatants stimulated with MERS S protein from immunized mice were measured, using ELISA. ( C ) IFN-γ, IL-2, and tumor necrosis factor α (TNF-α)-producing CD4+ T-cells in splenocytes were counted by flow cytometry. ( D ) IFN-γ, IL-2, and TNF-α producing polyfunctional CD4+ T-cells in splenocytes were analyzed by flow cytometry. *, p
    Interleukin 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Targeted disruption of SOCS3 promotes activation of BMP2-Smad signaling. (A) MC3T3/E1 cells were treated with BMP2 (100 ng/mL), and the phosphorylation level of Smad protein was examined by Western blotting. Shown are representative blots from three independent experiments. (B) SOCS3 knockdown and control cell lines were generated by lentivirus infection of MC3T3/E1 cells. The cells were then lysed and SOCS3 knockdown efficiency was examined by Western blotting. Shown are representative blots from three independent experiments. (C) SOCS3 knockdown and control cell lines were treated with BMP2 (100 ng/mL) for 0, 5, 15, 30 min and the phosphorylation level of Smad protein was detected by Western blotting. Shown are representative blots from three independent experiments. (D) BMSCs harvested from SOCS3 KD and WT mice were treated with BMP2 (100 ng/mL) or / and IL23 (100 ng/mL) cytokines. The cells were stimulated for 21 days and then stained with alizarin red S. Shown are representative photomicrographs.

    Journal: Frontiers in Immunology

    Article Title: Silencing SOCS3 Markedly Deteriorates Spondyloarthritis in Mice Induced by Minicircle DNA Expressing IL23

    doi: 10.3389/fimmu.2018.02641

    Figure Lengend Snippet: Targeted disruption of SOCS3 promotes activation of BMP2-Smad signaling. (A) MC3T3/E1 cells were treated with BMP2 (100 ng/mL), and the phosphorylation level of Smad protein was examined by Western blotting. Shown are representative blots from three independent experiments. (B) SOCS3 knockdown and control cell lines were generated by lentivirus infection of MC3T3/E1 cells. The cells were then lysed and SOCS3 knockdown efficiency was examined by Western blotting. Shown are representative blots from three independent experiments. (C) SOCS3 knockdown and control cell lines were treated with BMP2 (100 ng/mL) for 0, 5, 15, 30 min and the phosphorylation level of Smad protein was detected by Western blotting. Shown are representative blots from three independent experiments. (D) BMSCs harvested from SOCS3 KD and WT mice were treated with BMP2 (100 ng/mL) or / and IL23 (100 ng/mL) cytokines. The cells were stimulated for 21 days and then stained with alizarin red S. Shown are representative photomicrographs.

    Article Snippet: Recombinant IL23 and BMP2 were purchased from eBioscience.

    Techniques: Activation Assay, Western Blot, Generated, Infection, Mouse Assay, Staining

    IL23 expressed by minicircle DNA induces severe joint destruction and extensive bone loss in SOCS3 knockdown transgenic mice. (A–E) SOCS3 knockdown and WT control mice (20 mice per group, 10 male, and 10 female, 6–7 weeks old BALB/c mice were used in the experiment) were hydrodynamically injected with mc-IL23 or mc-Luc for 3 times at 1 month interval during the 4 months observation period. (A, B) The mice ankles and toe joints were examined and photographed (A) and the clinical arthritis scores were also recorded (B) when the animals were 5 months old. (C) Shown are representative micrographs of hematoxylin and eosin (HE) staining of ankle and toe joints sections from 5 months old mice. (D) Shown are the average clinical arthritis scores of joints observed from the HE staining in (C) . Data are representative of similar results from 6 mice. The error bars represent the ± S.D., ** P

    Journal: Frontiers in Immunology

    Article Title: Silencing SOCS3 Markedly Deteriorates Spondyloarthritis in Mice Induced by Minicircle DNA Expressing IL23

    doi: 10.3389/fimmu.2018.02641

    Figure Lengend Snippet: IL23 expressed by minicircle DNA induces severe joint destruction and extensive bone loss in SOCS3 knockdown transgenic mice. (A–E) SOCS3 knockdown and WT control mice (20 mice per group, 10 male, and 10 female, 6–7 weeks old BALB/c mice were used in the experiment) were hydrodynamically injected with mc-IL23 or mc-Luc for 3 times at 1 month interval during the 4 months observation period. (A, B) The mice ankles and toe joints were examined and photographed (A) and the clinical arthritis scores were also recorded (B) when the animals were 5 months old. (C) Shown are representative micrographs of hematoxylin and eosin (HE) staining of ankle and toe joints sections from 5 months old mice. (D) Shown are the average clinical arthritis scores of joints observed from the HE staining in (C) . Data are representative of similar results from 6 mice. The error bars represent the ± S.D., ** P

    Article Snippet: Recombinant IL23 and BMP2 were purchased from eBioscience.

    Techniques: Transgenic Assay, Mouse Assay, Injection, Staining

    Disruption of SOCS3 expression enhances the activation of STAT3 pathway by IL23. (A) BMSC cells were stimulated with IL23 (100 ng/mL) for 0, 5, 10, 15, 30, 60 min, followed by Western blotting to evaluate the activation of STAT3 and the expression of SOCS3. Shown are representative blots from three independent experiments. (B) The level of p-STAT3 in (A) were quantified by densitometry, and normalized to total STAT3 expression levels (the control set to 100). The error bars represent the ±S.D. from the mean. (C) BMSCs were isolated from SOCS3 KD and WT mice and stimulated with IL23 (100 ng/mL) for 0, 15, 30, 60 min, followed by Western blotting to detect the phosphorylation level of STAT3. Shown are representative blots from three independent experiments. (D) The level of p-STAT3 in (C) were quantified by densitometry, and normalized to total STAT3 expression levels (the WT control set to 100). Shown are representative data from three independent experiments. The error bars represent the ±S.D. from the mean, ** P

    Journal: Frontiers in Immunology

    Article Title: Silencing SOCS3 Markedly Deteriorates Spondyloarthritis in Mice Induced by Minicircle DNA Expressing IL23

    doi: 10.3389/fimmu.2018.02641

    Figure Lengend Snippet: Disruption of SOCS3 expression enhances the activation of STAT3 pathway by IL23. (A) BMSC cells were stimulated with IL23 (100 ng/mL) for 0, 5, 10, 15, 30, 60 min, followed by Western blotting to evaluate the activation of STAT3 and the expression of SOCS3. Shown are representative blots from three independent experiments. (B) The level of p-STAT3 in (A) were quantified by densitometry, and normalized to total STAT3 expression levels (the control set to 100). The error bars represent the ±S.D. from the mean. (C) BMSCs were isolated from SOCS3 KD and WT mice and stimulated with IL23 (100 ng/mL) for 0, 15, 30, 60 min, followed by Western blotting to detect the phosphorylation level of STAT3. Shown are representative blots from three independent experiments. (D) The level of p-STAT3 in (C) were quantified by densitometry, and normalized to total STAT3 expression levels (the WT control set to 100). Shown are representative data from three independent experiments. The error bars represent the ±S.D. from the mean, ** P

    Article Snippet: Recombinant IL23 and BMP2 were purchased from eBioscience.

    Techniques: Expressing, Activation Assay, Western Blot, Isolation, Mouse Assay

    Generation of IL23-induced SpA and SOCS3 knockdown transgenic mice. (A) 293T cells were transfected with equal moles of parental plasmids or its relevant minicircle DNA (mc-IL23). 48 h post-transfection, cells were lysed and the expression of IL23 was assessed by Western blotting using indicated antibodies. (B) Mc-IL23 or mc-Luc control was injected into the 6–7 weeks old female BALB/c mice for 3 times at 1 month interval by hydrodynamic method (6 mice per group). After indicated weeks post injection, serum samples were collected and evaluated by ELISA to detect IL23 protein level. The error bars represent the ±S.D. from the mean, ** P

    Journal: Frontiers in Immunology

    Article Title: Silencing SOCS3 Markedly Deteriorates Spondyloarthritis in Mice Induced by Minicircle DNA Expressing IL23

    doi: 10.3389/fimmu.2018.02641

    Figure Lengend Snippet: Generation of IL23-induced SpA and SOCS3 knockdown transgenic mice. (A) 293T cells were transfected with equal moles of parental plasmids or its relevant minicircle DNA (mc-IL23). 48 h post-transfection, cells were lysed and the expression of IL23 was assessed by Western blotting using indicated antibodies. (B) Mc-IL23 or mc-Luc control was injected into the 6–7 weeks old female BALB/c mice for 3 times at 1 month interval by hydrodynamic method (6 mice per group). After indicated weeks post injection, serum samples were collected and evaluated by ELISA to detect IL23 protein level. The error bars represent the ±S.D. from the mean, ** P

    Article Snippet: Recombinant IL23 and BMP2 were purchased from eBioscience.

    Techniques: Transgenic Assay, Mouse Assay, Transfection, Expressing, Western Blot, Injection, Enzyme-linked Immunosorbent Assay

    Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, TNF-α, IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p

    Journal: Frontiers in Immunology

    Article Title: The IDO–AhR Axis Controls Th17/Treg Immunity in a Pulmonary Model of Fungal Infection

    doi: 10.3389/fimmu.2017.00880

    Figure Lengend Snippet: Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, TNF-α, IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p

    Article Snippet: The levels of IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17, IL-22, IL-27, IL-35, IL-23, TNF-α, IFN-γ, and TGF-β were measured by capture enzyme-linked immunosorbent assay (ELISA) with antibody pairs purchased from eBioscience or PBL.

    Techniques: Expressing, Mouse Assay, Infection, Real-time Polymerase Chain Reaction

    Indoleamine 2,3 dioxygenase-1 (IDO1) expression controls the presence of intracellular cytokines in pulmonary CD11b + and CD11c + cells. The intracellular cytokines were determined in CD11b + (A) and CD11c + (B) lung infiltrating leukocytes of wild-type and IDO1 −/− C57BL/6 mice after 96 h, 2, and 10 weeks of infection with 1 × 10 6 Paracoccidioides brasiliensis yeasts. The lung cells were obtained as described in Section “ Materials and Methods ” and labeled with antibodies conjugated to different fluorochromes. The lung infiltrating leukocytes were gated by FSC/SSC analysis. The cells were gated for CD11b + or CD11c + expression and then for the presence of cytokines IL-12, TNF-α, IL-1β, IL-10, IL-6, and TGF-β. One hundred thousand cells were acquired on FACS CANTO II and subsequently analyzed by FlowJo software. Data are expressed as means ± SEM and are representative of three independent experiments using five mice of each mouse strain per group (* p

    Journal: Frontiers in Immunology

    Article Title: The IDO–AhR Axis Controls Th17/Treg Immunity in a Pulmonary Model of Fungal Infection

    doi: 10.3389/fimmu.2017.00880

    Figure Lengend Snippet: Indoleamine 2,3 dioxygenase-1 (IDO1) expression controls the presence of intracellular cytokines in pulmonary CD11b + and CD11c + cells. The intracellular cytokines were determined in CD11b + (A) and CD11c + (B) lung infiltrating leukocytes of wild-type and IDO1 −/− C57BL/6 mice after 96 h, 2, and 10 weeks of infection with 1 × 10 6 Paracoccidioides brasiliensis yeasts. The lung cells were obtained as described in Section “ Materials and Methods ” and labeled with antibodies conjugated to different fluorochromes. The lung infiltrating leukocytes were gated by FSC/SSC analysis. The cells were gated for CD11b + or CD11c + expression and then for the presence of cytokines IL-12, TNF-α, IL-1β, IL-10, IL-6, and TGF-β. One hundred thousand cells were acquired on FACS CANTO II and subsequently analyzed by FlowJo software. Data are expressed as means ± SEM and are representative of three independent experiments using five mice of each mouse strain per group (* p

    Article Snippet: The levels of IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17, IL-22, IL-27, IL-35, IL-23, TNF-α, IFN-γ, and TGF-β were measured by capture enzyme-linked immunosorbent assay (ELISA) with antibody pairs purchased from eBioscience or PBL.

    Techniques: Expressing, Mouse Assay, Infection, Labeling, FACS, Software

    Role played by interleukin (IL)–23. Groups of mice were intranasally inoculated with 2 × 10 6 yeast cells, and lungs were analyzed at serial intervals for expression of IL-23 by quantitative reverse-transcription polymerase chain reaction

    Journal:

    Article Title: Interleukins 17 and 23 Influence the Host Response to Histoplasma capsulatum

    doi: 10.1086/599333

    Figure Lengend Snippet: Role played by interleukin (IL)–23. Groups of mice were intranasally inoculated with 2 × 10 6 yeast cells, and lungs were analyzed at serial intervals for expression of IL-23 by quantitative reverse-transcription polymerase chain reaction

    Article Snippet: qRT-PCR for IL-17A, IL-17F, and IL-23 was performed using TaqMan primers (Applied Biosystems).

    Techniques: Mouse Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    Analysis of T-cell activation after immunization with the RNA adjuvant and MERS S protein vaccine. ( A ) The population of MERS S protein-specific cells secreting interferon-γ (IFN-γ) and interleukin-2 (IL-2) were quantified by using ELISPOT assay, after treatment with MERS S protein in cultured splenocytes from immunized mice. ( B ) Cytokine levels in splenocyte supernatants stimulated with MERS S protein from immunized mice were measured, using ELISA. ( C ) IFN-γ, IL-2, and tumor necrosis factor α (TNF-α)-producing CD4+ T-cells in splenocytes were counted by flow cytometry. ( D ) IFN-γ, IL-2, and TNF-α producing polyfunctional CD4+ T-cells in splenocytes were analyzed by flow cytometry. *, p

    Journal: Pharmaceutics

    Article Title: MERS-CoV Spike Protein Vaccine and Inactivated Influenza Vaccine Formulated with Single Strand RNA Adjuvant Induce T-Cell Activation through Intranasal Immunization in Mice

    doi: 10.3390/pharmaceutics12050441

    Figure Lengend Snippet: Analysis of T-cell activation after immunization with the RNA adjuvant and MERS S protein vaccine. ( A ) The population of MERS S protein-specific cells secreting interferon-γ (IFN-γ) and interleukin-2 (IL-2) were quantified by using ELISPOT assay, after treatment with MERS S protein in cultured splenocytes from immunized mice. ( B ) Cytokine levels in splenocyte supernatants stimulated with MERS S protein from immunized mice were measured, using ELISA. ( C ) IFN-γ, IL-2, and tumor necrosis factor α (TNF-α)-producing CD4+ T-cells in splenocytes were counted by flow cytometry. ( D ) IFN-γ, IL-2, and TNF-α producing polyfunctional CD4+ T-cells in splenocytes were analyzed by flow cytometry. *, p

    Article Snippet: The concentrations of interferon γ (IFN-γ), interleukin-2 (IL-2), IL-6, and tumor necrosis factor α (TNF-α) were detected with ELISA kits (Invitrogen; Thermo Fisher Scientific Inc., Waltham, MA, USA), according to the manufacturer’s instructions.

    Techniques: Activation Assay, Enzyme-linked Immunospot, Cell Culture, Mouse Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry