Journal: bioRxiv

Article Title: Vasoactive intestinal peptide confers anticipatory mucosal immunity by regulating ILC3 activity

doi: 10.1101/729400

Figure Lengend Snippet: a - c , Mixed bone marrow chimeras were generated by reconstituting CD45.1 +/+ lethally irradiated recipients with F1 (CD45.1 × CD45.2, CD45.1 + CD45.2 + ) wild-type and CD45.2 +/+ Bmal1 ΔIL-7R bone marrow. a , Representative contour plots of frequency of different lymphocyte subsets isolated in the small intestine of mixed bone marrow chimeras. Plots show live CD45.1 + CD45.2 + cells for wild-type (F1) mice (upper panels) or CD45.2 +/+ cells for Bmal1 -deficient cells (lower panels). b , Frequency of different lymphocyte subsets derived from wild-type (red) and Bmal1 -deficient (blue) donor bone marrow isolated from the small intestine of chimeric mice. c , Frequency of wild-type (CD45.1 + CD45.2 + , red) and Bmal1 -deficient (CD45.2 +/+ , blue) IL-22-producing ILC3 isolated from the small intestine of chimeric mice. a - c Data are representive of 2 independent experiments ( a ) and show the mean ± s.e.m. for one experiment ( n = 6-8 mice/experiment) ( b , c ). d , 4 × 10 3 ILC3 were purified from the small intestine of wild-type (red) or Bmal1 ΔIL7R (blue) mice and cultivated 16 hours in presence of recombinant IL-7 (10ng/mL) to determine constitutive IL-22 expression, or IL-7 together with IL-23 (10 ng/mL) to elicit maximal IL-22 cytokine expression. IL-22 production was determined in culture supernatants by ELISA. e-f , Frequency of IL-5 and IL-22-producing ( e ) lung ILC2, and ( f ) small intestinal ILC3 at T4 and T16 isolated from wild-type and Bmal1 ΔIL7R mice. Left panel shows the constitutive expression of cytokines; right panel shows the cytokine expression after cultivation with IL-7 (10 ng/mL) and IL-23 (10 ng/mL). e , f Data show the mean ± s.e.m. of one of two similar experiments ( n = 4 mice for each genotype per time point per experiment).

Article Snippet: For longer term cultures, media was supplemented with 10 ng/mL recombinant human IL-7 (R&D Systems)(a survival factor), with or without 10ng/mL recombinant murine IL-23 (R&D Systems).

Techniques: Generated, Irradiation, Isolation, Derivative Assay, Purification, Recombinant, Expressing, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Vasoactive intestinal peptide confers anticipatory mucosal immunity by regulating ILC3 activity

doi: 10.1101/729400

Figure Lengend Snippet: a , Food consumption of mice was determined at the indicated time points. Data show the mean ± s.e.m for one of two similar experiments ( n = 12 mice per experiment). b , Schematic representation of the time-restricted feeding regime used in c and d . Mice were given food only during the day (day food) or the night (night food) for seven days. All mice were analysed at T4 on day 8. c,d , Analyses of IL-5-expressing ILC2 ( c ) and IL-22-expressing ILC3 ( d ) from the small intestine, mesenteric LN and lung of mice fed only during the day (white) or only during the night (blue) as determined by flow cytometric analyses of intracellular cytokine. Representiative flow cytometric histograms of cytokine expression (left panels) and the frequency of constitutively producing IL-5 + or IL-22 + ILC2 and ILC3, respectively (right panels), showing individual mice (circles) together with the mean ± s.e.m. pooled from two independent experiments (gut and mLN: n = 6, 6 mice for each condition/experiment; lung: n = 4, 6 mice for each condition/experiment). e , Mice were treated with combination antibiotics (ampicillin, streptomycin and colistin) for two weeks before analysis of constitutive IL-22 production by intracellular staining. Data show the mean ± s.e.m ( n =6 mice per experimental condition). f , g , Expression of IL-5-expressing ILC2 ( f ) and IL-22-expressing ILC3 ( g ) from the small intestine and mesenteric LN from mice fed (green) or fasted for 16 hours (white). f , g Data show the mean ± s.e.m. of ( n = 6 mice per group per condition) and show one of two similar experiments. h , Gut ILC3 (7.5 × 10 3 ) were purified from fed (green) or fasted for 14 h (white) and cultivated 16 hours in presence of IL-7 (10 ng/mL) alone (constitutive expression) and IL-7 and IL-23 (10 ng/mL). IL-22 secretion was determined in culture supernatants by ELISA. Data show the response of cells pooled from 2 independent experiments with 3 mice per well for each condition in each experiment.

Article Snippet: For longer term cultures, media was supplemented with 10 ng/mL recombinant human IL-7 (R&D Systems)(a survival factor), with or without 10ng/mL recombinant murine IL-23 (R&D Systems).

Techniques: Expressing, Staining, Purification, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Vasoactive intestinal peptide confers anticipatory mucosal immunity by regulating ILC3 activity

doi: 10.1101/729400

Figure Lengend Snippet: a , Intracellular expression of IL-22 detected in ILC3 after four hours of incubation with IL-7 and IL-23 (10 ng/mL) in presence of vehicle (DMSO), VIP (1 μM) or Dibutyryl cyclic-AMP (10 μM). b , IL-22 secretion by purified ILC3 isolated from the small intestine and cultivated for 16 hours with IL-7 (10 ng/mL) or IL-7 and IL-23 (10 ng/mL) in presence of vehicle (DMSO) or VIP (1 μM). IL-22 was measured in the supernatant by ELISA. Data are a pool of three independent experiments (mean ± s.e.m) from a total of six experiments. c , d , Titration effect of VIP ( c ) and Dibutyryl cyclic-AMP ( d ) on the production of IL-22 by intestinal ILC3. Purified ILC3 were stimulated with IL-7 alone or IL-7 and IL-23 (10 ng/mL) and different concentrations of either VIP ( c ) or dibutyryl cyclic-AMP ( d ). ( e ) IL-22 production of ILC3 cultured with IL-7 (10 ng/mL) or IL-7 and IL-23 (10 ng/mL) in the presence of VIP (100mM) or VIPR2 agonist BAY55-9837 (100mM). f, IL-22 production of ILC3 cultured with IL-7 and IL-23 (10 ng/mL) in the presence of VIP (100mM) or VIP (100mM) with the VIPR2 antagonist PG99-465 (1μM) for 16 hours. e , f , Data are pooled from two independent experiments, each dot represents a pool of two mice (mean ± s.e.m).

Article Snippet: For longer term cultures, media was supplemented with 10 ng/mL recombinant human IL-7 (R&D Systems)(a survival factor), with or without 10ng/mL recombinant murine IL-23 (R&D Systems).

Techniques: Expressing, Incubation, Purification, Isolation, Enzyme-linked Immunosorbent Assay, Titration, Cell Culture

Journal: bioRxiv

Article Title: Vasoactive intestinal peptide confers anticipatory mucosal immunity by regulating ILC3 activity

doi: 10.1101/729400

Figure Lengend Snippet: a , Mice were injected i.p. with 2 nM VIP or PBS 2h (T2) before sacrifice (T4). Constitutive expression of IL-5 by ILC2, and IL-22 and IL-17 by ILC3 from the small intestine was determined by flow cytometry. Data show the mean ± s.e.m. of one of two similar experiments ( n = 5-6 mice per time point per experiment). b , Intracellular expression of ILC3 from the small intestine of wild-type or Vipr2 −/− mice after 4 hours of incubation with IL-7. Data show the mean ± s.e.m. of one of two similar experiments ( n = 8 mice per time point per experiment). c , 4.5 × 10 3 ILC3 purified from small intestine of wild-type or Vipr2 −/− were cultivated with IL-7 (10 ng/mL) alone or IL-7 and IL-23 (10 ng/mL) in presence of vehicle or VIP (1 μM) for 16 hours. IL-22 was measured in the supernatant by ELISA. Data represent a pool of 2 independent experiments and each dot represents a pool of two mice (mean ± s.e.m).

Article Snippet: For longer term cultures, media was supplemented with 10 ng/mL recombinant human IL-7 (R&D Systems)(a survival factor), with or without 10ng/mL recombinant murine IL-23 (R&D Systems).

Techniques: Injection, Expressing, Flow Cytometry, Incubation, Purification, Enzyme-linked Immunosorbent Assay

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 1. TH17 cells derived in vitro show different metabolic states. (A) RT-PCR analysis of key glycolytic pathway genes in TH17 (b) and TH17 (23) cells differentiated in vitro for 48 h; gene expression was normalized to Actb. (B) ECAR of TH17 (b) and TH17 (23) cells differentiated for 96 h assessed by a glycolytic stress test. ECAR was measured under basal conditions and in response to glucose (10 mM), oligomycin (1.0 mM), and 2-deoxyglucose (2-DG) (50 mM). (C) Principal component analysis (PCA) analysis of identified metabolites of TH17 (b) and TH17 (23) cells (n = 5) differentiated in vitro for 48 h by metabolomics. (D) Metabolomics analysis of TH17 (b) and TH17 (23) cells differentiated in vitro for 48 h; the top differentially observed metabolites are shown in heat map. Data are representative of three independent experiments (A and B). Error bars represent SEM. *p , 0.05, **p , 0.01, ***p , 0.001, determined by two-tailed unpaired t test.

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: Derivative Assay, In Vitro, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Two Tailed Test

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 2. TH17 cells show differential metabolic pathway gene activation in vivo. (A) Scatter plot shows differential expressed genes between CNS TH17 (red) and ileum TH17 (blue) cells with differential TH17 signature genes highlighted (black dot). differentially-expressed genes are filtered as false discovery rate (FDR) , 0.05 and FC $ 1.5 from DESeq2. (B) Scatter plot shows differential expressed genes between CNS TH17 (red) and ileum TH17 (blue) cells with glycolysis pathway genes highlighted (black dot) and differential genes labeled. DEGs are filtered as in (A). (C) Scatter plot shows differential expressed genes between CNS TH17 (red) and ileum TH17 (blue) cells with glutaminolysis pathway genes highlighted (black dot) and differential genes labeled. DEGs are filtered as in (A). (D) CNS TH17 and ileum TH17 cells depend on distinct metabolic pathways. Numbers indicate the differentially expressed genes and total genes for each selected pathway. (E) KEGG pathway enrichment for CNS TH17 cell highly expressed genes. (F) KEGG pathway enrichment for ileum TH17 cell highly expressed genes. Color represents log-transferred FDR, and dot size represents the number of differentially expressed genes observed for this pathway (E and F).

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: Activation Assay, In Vivo, Labeling

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 3. TH17 cells derived in vitro show discrete chromatin states. (A) Scatter plot for differentially opened/closed regions (OCRs) between TH17 (23) (red) and TH17 (b) cells (blue) with differential numbers labeled. (B) ATAC-seq reads density heatmap around the OCRs (shows the peak summit 6 1 kb region) between TH17 (23) and TH17 (b) cells. (C) The enriched transcription factor motifs identified from top 1000 differentially OCRs between TH17 (23) cells and TH17 (b) cells; color bar represents log-transferred p values. Right panel shows the identified motif sequences for key regulators. (D) WashU Epigenome Browser visualization of differentially OCR signals for key glycolytic genes in TH17 (23) and TH17 (b) cells. Arrows represent the NF-kB binding motif sites.

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: Derivative Assay, In Vitro, Labeling, Binding Assay

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 4. TH17 cells show discrete chromatin states in vivo. (A) Scatter plot for differentially opened/closed regions (OCRs) between CNS TH17 (red) and ileum TH17 cells (blue) with differential numbers labeled. (B) ATAC-seq reads density heatmap around the OCRs (shows the peak summit 6 1 kb region) between CNS TH17 and ileum TH17 cells. (C) The enriched transcription factor motifs identified from top 1000 differential OCRs between CNS TH17 cells and ileum TH17 cells; color bar represents log-transferred p values. Right panel shows the identified motif sequences for key regulators. (D) WashU Epigenome Browser visualization of differentially OCR signals with corresponding gene expression levels for key TH17 signature genes and metabolic genes in CNS TH17 and ileum TH17 cells. Arrows represent the NF-kB binding motif sites.

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: In Vivo, Labeling, Gene Expression, Binding Assay

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 5. Identification of a miR-21–Peli1–c-Rel pathway controlling pathogenic TH17 cell glycolysis. (A) WashU Epigenome Browser visualization of ATAC-seq signals across selected microRNA genome loci in ileum homeostatic and CNS-infiltrated pathogenic TH17 cells. (B) Relative expression of microRNA in ileum homeostatic and CNS-infiltrated pathogenic TH17 cells (n = 3). (C) KEGG pathway enrichment for genes downregulated in miR-212/2

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: Expressing

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

Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.

doi: 10.4049/jimmunol.2000060

Figure Lengend Snippet: FIGURE 6. Pharmaceutical inhibition c-Rel–mediated glycolysis in pathogenic TH17 cells prevents autoimmunity. (A) Western blot results for c-Rel and RelA from TH17 cells differentiated under indicated conditions. (B) Western blot results for c-Rel and RelA from TH17 cells differentiated under indicated conditions. (C) Western blot results for c-Rel and RelA from TH17 cells differentiated under indicated conditions. (D) Normalized RT-PCR results for key glycolytic pathway genes in TH17 (23) cells differentiated in vitro for 48 h, treated with H2O or 500 mg/ml PTXF for 16 h. (E) ECAR of TH17 (23) cells differentiated for 48 h and then were treated with H2O or 500 mg/ml PTXF for 16 h, assessed by a glycolysis stress test. (F) Basal and maximal glycolytic capability of TH17 (23) cells differentiated for 48 h and then were treated with H2O or PTXF for 16 h. (G) EAE development in C57BL/6 mice, i.p. PBS or 100 mg/kg PTXF per mouse (n = 6) from day 7 to day 14. (H) EAE development in recipient C57BL/6 mice (sublethal irradiation; n = 6) i.p. with pathogenic TH17 (23) cells differentiated from naive 2D2 CD4+ T cells for 96 h and then treated with H2O or 500 mg/ml PTXF for 16 h. Data shown are one experiment representative of three independent experiments (A–C, G, and H). Data are representative of three independent experiments (D–F). Error bars represent SEM. *p , 0.05, **p , 0.01, ***p , 0.001, determined by two-tailed unpaired t test.

Article Snippet: Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).

Techniques: Inhibition, Western Blot, Reverse Transcription Polymerase Chain Reaction, In Vitro, Irradiation, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity

doi: 10.1016/j.xcrm.2022.100744

Figure Lengend Snippet:

Article Snippet: Rec. IL-23 , R & D System , Cat# 1887-ML/CF.

Techniques: In Vivo, Control, Recombinant, In Situ, Enzyme-linked Immunosorbent Assay, Extraction, Cytotoxicity Assay, Clinical Proteomics, Gene Expression, Software