Journal: Frontiers in Immunology

Article Title: IL-17A-associated PTGS2 and MMP9 inflammatory signaling in ischemic stroke: clinical correlation and experimental evidence

doi: 10.3389/fimmu.2026.1812571

Figure Lengend Snippet: Bioinformatics analysis prioritized PTGS2 and MMP9 as candidate inflammatory genes associated with the IL-17 signaling pathway in ischemic stroke. (A) Venn diagram showing the overlap between differentially expressed genes (DEGs) identified from GEO datasets and genes retrieved from the CellAge database. Five overlapping genes were identified: PTGS2, MMP9, SERPINB2, CTNNAL1, and HTRA1. CS, genes from CellAge database; DEG, significantly differentially expressed genes in ischemic stroke. (B) Gene Ontology (GO) enrichment analysis of the overlapping genes, including biological process, cellular component, and molecular function categories. (C) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showing that the overlapping genes were enriched in inflammation-related pathways, including the IL-17 signaling pathway and TNF signaling pathway. (D) GSEA comparing ischemic stroke (IS) and control samples, demonstrating enrichment of immune- and inflammation-related pathways in IS. (E) Predicted transcription factor–target gene regulatory network for PTGS2 and MMP9, showing candidate shared upstream transcription factors. Yellow nodes represent TFs, red nodes represent target genes.

Article Snippet: Rats in the IL-17A neutralizing antibody-treated group were treated with Anti-Mouse/Rat IL-17A Antibody (17F3; MedChemExpress, Cat. No. HY-P990222), a mouse IgG1κ monoclonal neutralizing antibody against mouse/rat IL-17A, via intravenous injection at 100 μg/rat immediately after reperfusion and again at 12 h post-reperfusion.

Techniques: Control

Journal: Frontiers in Immunology

Article Title: IL-17A-associated PTGS2 and MMP9 inflammatory signaling in ischemic stroke: clinical correlation and experimental evidence

doi: 10.3389/fimmu.2026.1812571

Figure Lengend Snippet: Serum IL-17A, PTGS2, and MMP9 levels in the clinical cohort and their correlations with each other and with stroke severity. (A–C) Serum IL-17A, PTGS2, and MMP9 levels were significantly higher in patients with ischemic stroke than in healthy controls. IS, ischemic stroke (n = 104); Normal, control group (n = 58). (D) Correlations among serum IL-17A, PTGS2, and MMP9 levels in the stroke group. (E) Positive correlations between serum IL-17A, PTGS2, and MMP9 levels and admission NIHSS scores in patients with ischemic stroke. Correlations were analyzed using Spearman’s rank correlation test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Rats in the IL-17A neutralizing antibody-treated group were treated with Anti-Mouse/Rat IL-17A Antibody (17F3; MedChemExpress, Cat. No. HY-P990222), a mouse IgG1κ monoclonal neutralizing antibody against mouse/rat IL-17A, via intravenous injection at 100 μg/rat immediately after reperfusion and again at 12 h post-reperfusion.

Techniques: Control

Journal: Frontiers in Immunology

Article Title: IL-17A-associated PTGS2 and MMP9 inflammatory signaling in ischemic stroke: clinical correlation and experimental evidence

doi: 10.3389/fimmu.2026.1812571

Figure Lengend Snippet: MCAO-induced activation of IL-17A/PTGS2/MMP9-related inflammatory markers and changes after anti-IL-17A antibody treatment in rat brain tissue. (A–C) Relative mRNA levels of PTGS2, MMP9, and IL-17A in the sham, MCAO, and anti-IL-17A Ab groups. (D–F) Protein concentrations of PTGS2, MMP9, and IL-17A. (G, H) Concentrations of PGE2 and IL-6. (I) Neurological deficit scores assessed using the Longa scoring system. In panel I, the sham group median was 0; therefore, a minimal bar height was displayed for visualization only. (J) Representative TTC-stained brain sections from the MCAO and anti-IL-17A Ab groups. White areas indicate infarct regions, whereas red areas indicate viable brain tissue. (K) Quantitative analysis of infarct volume percentage based on TTC staining. Sham, sham-operated group; Model, MCAO group; anti-IL-17A Ab, IL-17A neutralizing antibody-treated MCAO group. Data in panels A-H are presented as mean ± SEM (n = 6 per group). Neurological deficit scores in panel I are presented as median (interquartile range) (n = 6 per group). TTC analysis in panels (J, K) was performed in an independent cohort of rats, with final sample sizes of n = 4 for the MCAO group and n = 5 for the anti-IL-17A Ab group; one rat in the MCAO group died before tissue collection. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Rats in the IL-17A neutralizing antibody-treated group were treated with Anti-Mouse/Rat IL-17A Antibody (17F3; MedChemExpress, Cat. No. HY-P990222), a mouse IgG1κ monoclonal neutralizing antibody against mouse/rat IL-17A, via intravenous injection at 100 μg/rat immediately after reperfusion and again at 12 h post-reperfusion.

Techniques: Activation Assay, Staining

Journal: Frontiers in Immunology

Article Title: IL-17A-associated PTGS2 and MMP9 inflammatory signaling in ischemic stroke: clinical correlation and experimental evidence

doi: 10.3389/fimmu.2026.1812571

Figure Lengend Snippet: Spearman correlation analyses among IL-17A, PTGS2, MMP9, PGE2, and IL-6 in rat brain tissue. Correlation analyses were performed using pooled data from all animals (n = 18). (A) Correlation between IL-17A and PTGS2 mRNA expression. (B) Correlation between IL-17A and MMP9 mRNA expression. (C) Correlation between IL-17A and PTGS2 protein expression. (D) Correlation between IL-17A and MMP9 protein expression. (E) Correlation between IL-17A and PGE2 levels. (F) Correlation between IL-17A and IL-6 levels. Sham, sham-operated group; Model, MCAO group; anti-IL-17A Ab, IL-17A neutralizing antibody-treated MCAO group. Given the limited sample size, these correlations should be interpreted with caution.

Article Snippet: Rats in the IL-17A neutralizing antibody-treated group were treated with Anti-Mouse/Rat IL-17A Antibody (17F3; MedChemExpress, Cat. No. HY-P990222), a mouse IgG1κ monoclonal neutralizing antibody against mouse/rat IL-17A, via intravenous injection at 100 μg/rat immediately after reperfusion and again at 12 h post-reperfusion.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Attenuates Systemic Inflammation and Downregulates Th17-Related Factor Expression. ( A ) KEGG pathway enrichment analysis of downstream targets of OST. This figure is based on data from KEGG (Kyoto Encyclopedia of Genes and Genomes) at https://www.kegg.jp/ ; © Kanehisa Laboratories. ( B ) Flow cytometry was used to evaluate the proportion of CD4⁺IL-17A⁺ cells in the spleen. ( C ) Immunohistochemical staining of liver tissues was performed to assess IL-17A expression. ( D ) Immunofluorescence staining was conducted to detect CD4⁺RORγt⁺ double-positive cells in liver sections. ( E ) Serum levels of IL-6, IL-1β, TNF-α, and IL-17A were measured using ELISA. n = 6. **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Expressing, Flow Cytometry, Immunohistochemical staining, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Inhibits Th17 Cell Differentiation in Vitro. ( A ) Flow cytometry was used to assess the proportion of CD4⁺IL-17A⁺ cells in a Th17-polarized cell model treated with different doses of OST. ( B ) RT-qPCR was performed to evaluate mRNA expression levels of RORγt and IL-17A. ( C ) Western blot was performed to evaluate protein expression levels of RORγt, IL-17A, IL-17RA, TRAF6, and Act1. ( D ) ELISA was used to quantify the secretion of IL-6, IL-1β, TNF-α, IL-10, and TGF-β in the culture supernatants. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Suppresses Th17 Differentiation via an ADRA1D-Dependent Mechanism. ( A ) Volcano plot of differentially expressed genes (DEGs) in the GSE110729 obesity dataset. ( B ) Venn diagram showing the intersection between OST’s potential targets and DEGs from the GSE110729 dataset. ( C ) RT-qPCR was performed to assess the expression of ADRA1D, IL-17A, and RORγt in Th17-polarized CD4⁺ T cells with or without ADRA1D overexpression. ( D ) Western blot was conducted to evaluate the protein expression of ADRA1D, IL-17A, RORγt, IL-17RA, TRAF6, and Act1. ( E ) ELISA was used to measure the levels of IL-6, IL-1β, TNF-α, IL-10, and TGF-β in the culture supernatants. n = 3. *** P < 0.001, **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Quantitative RT-PCR, Expressing, Over Expression, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: ADRA1D Overexpression Attenuates the Metabolic and Immunomodulatory Effects of OST in Vivo. ( A ) Body weight changes were monitored in HFD-fed mice treated with OST. (B) Weights of adipose tissue and liver were measured to assess the effect on organ hypertrophy. ( C ) Serum levels of TG, TC, FFA, ALT, and AST were evaluated by biochemical assays. ( D ) Flow cytometry was performed to assess the proportion of CD4⁺IL-17A⁺ cells in the spleen. ( E ) Immunofluorescence staining of liver tissue was used to detect CD4⁺RORγt⁺ double-positive cell infiltration. ( F ) Immunohistochemistry was performed to evaluate hepatic IL-17A protein expression. ( G ) Western blotting was conducted to assess ADRA1D protein expression in liver tissue. ( H ) Western blot was used to detect the phosphorylation levels of ERK1/2 and PI3K in liver tissue. ( I ) H&E staining of adipose tissue was used to assess adipocyte morphology. (J) Oil red O staining of liver sections was performed to evaluate hepatic lipid deposition. n = 6. ** P < 0.01, **** P < 0.0001. For three or more groups, one-way or two-way analysis of variance (ANOVA) was applied, followed by Tukey’s post hoc test. P < 0.05 was considered statistically significant.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Over Expression, In Vivo, Flow Cytometry, Immunofluorescence, Staining, Immunohistochemistry, Expressing, Western Blot, Phospho-proteomics

Journal: Biomolecules & Therapeutics

Article Title: DW2007 Ameliorates Colitis and Rheumatoid Arthritis in Mice by Correcting Th17/Treg Imbalance and Inhibiting NF-κB Activation

doi: 10.4062/biomolther.2016.018

Figure Lengend Snippet: Effect of DW2007 on Th17 and Treg cell differentiation. (A) Effect on Th17 and Treg cell differentiation. Splenocytes were stimulated with anti-CD3 (1 μg/mL) and anti-CD28 (1 μg/mL) with or without IL-6 (20 ng/mL) plus TGF-β (1 ng/mL) in the presence of DW2007 (AC) for 5 days. Staining of cell surface CD4 antigen and intracellular IL-17 was performed and analyzed using flow cytometry. (B) Effect on RORγt, Foxp3, IL-17, and IL-10 expression by qRT-PCR. (C) Effect on IL-17 and IL-10 expression by ELISA. All data indicate mean ± SD (n=6). # p <0.05 vs. normal control. * p <0.05 vs. cells stimulated with anti-CD3/anti-CD28 and IL-6/TGFβ.

Article Snippet: The reaction mixture were filtered and T cells were isolated using a Pan T cell Isolation Kit II, fixed, stained with anti-Foxp3 or anti-IL-17A antibodies (Biogems, CA, USA) and analyzed by flow cytometry.

Techniques: Cell Differentiation, Staining, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Biomolecules & Therapeutics

Article Title: DW2007 Ameliorates Colitis and Rheumatoid Arthritis in Mice by Correcting Th17/Treg Imbalance and Inhibiting NF-κB Activation

doi: 10.4062/biomolther.2016.018

Figure Lengend Snippet: Effect of DW2007 on the differentiation to Th17 and Treg cells in wild-type mice with DSS-induced colitis. (A) Effect on Th17 (CD4 + IL17 + ) cell differentiation. (B) Effect on Treg (CD4 + Foxp3 + ) cell differentiation. DSS, except in the control group, was orally administered to mice treated with saline, DW2007 (AC), or sulfasalazine (SS). Test agents (AC10, 10 mg/kg DW2007; AC20, 20 mg/kg DW2007; or SS50, 50 mg/kg SS) or saline were orally administered for 3 days after DSS treatment. The mice were killed 18 h after the final administration of test agents. The CD4 + T cells were isolated from the lamina propria of the colon using Pan T cell isolation kit. So, we performed single staining for flow cytometry (Th17 cells: IL-17, Treg cells: Foxp3). Th17 and Treg cells were then analyzed by flow cytometry. All values are mean ± SD (n=6). # p <0.05 vs. control group. * p <0.05 vs. DSS group.

Article Snippet: The reaction mixture were filtered and T cells were isolated using a Pan T cell Isolation Kit II, fixed, stained with anti-Foxp3 or anti-IL-17A antibodies (Biogems, CA, USA) and analyzed by flow cytometry.

Techniques: Cell Differentiation, Control, Saline, Isolation, Cell Isolation, Staining, Flow Cytometry

Journal: Biomolecules & Therapeutics

Article Title: DW2007 Ameliorates Colitis and Rheumatoid Arthritis in Mice by Correcting Th17/Treg Imbalance and Inhibiting NF-κB Activation

doi: 10.4062/biomolther.2016.018

Figure Lengend Snippet: Effects of DW2007 on the expression of iNOS and COX-2, activation of NF-κB (A), and expression of inflammatory cytokines (B) and in TLR4 −/− mice with DSS-induced colitis. DSS, except in the normal control group (NOR), was orally administered to mice treated with saline (DSS), DW2007 (AC), or sulfasalazine (SS). Test agents (AC10, 10 mg/kg DW2007; AC20, 20 mg/kg AC; or SS50, 50 mg/kg SS) or saline were orally administered for 3 days after DSS treatment. iNOS, COX-2, and NF-κB were determined by immunoblotting. TNF-α, IL-1β, IL-6, IL-10, and IL-17 were determined by ELISA. All values are mean ± SD (n=6). # p <0.05 vs. control group. * p <0.05 vs. DSS group.

Article Snippet: The reaction mixture were filtered and T cells were isolated using a Pan T cell Isolation Kit II, fixed, stained with anti-Foxp3 or anti-IL-17A antibodies (Biogems, CA, USA) and analyzed by flow cytometry.

Techniques: Expressing, Activation Assay, Control, Saline, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules & Therapeutics

Article Title: DW2007 Ameliorates Colitis and Rheumatoid Arthritis in Mice by Correcting Th17/Treg Imbalance and Inhibiting NF-κB Activation

doi: 10.4062/biomolther.2016.018

Figure Lengend Snippet: Effect of DW2007 on the expression of T cell differentiation (A) and its cytokines (B) in CIA mice. Arthritis was induced by the first intradermal injection of bovine type II collagen and the second intradermal injection of collagen with complete Freund’s adjuvant at the base of the tail of DBA/1 J mice. Test agents (AC, 20 mg/kg DW2007; IB, 50 mg/kg ibuprofen; or CIA, vehicle alone) were orally administrated daily for 20 days from day 1 after the second injection. T cells were isolated from the spleen. Th17 and Treg cells were then analyzed by a flow cytometer. Cytokines were determined in the paw joint tissues by real-time PCR (for IL-10, IL-17a, IFN-γ, Foxp3, and β-actin). All data indicate mean ± SD (n=7). # p <0.05 vs. control group. * p <0.05 vs. CIA group.

Article Snippet: The reaction mixture were filtered and T cells were isolated using a Pan T cell Isolation Kit II, fixed, stained with anti-Foxp3 or anti-IL-17A antibodies (Biogems, CA, USA) and analyzed by flow cytometry.

Techniques: Expressing, Cell Differentiation, Injection, Adjuvant, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction, Control

Journal: Biomolecules & Therapeutics

Article Title: DW2007 Ameliorates Colitis and Rheumatoid Arthritis in Mice by Correcting Th17/Treg Imbalance and Inhibiting NF-κB Activation

doi: 10.4062/biomolther.2016.018

Figure Lengend Snippet: Effect of DW2007 on the expression of iNOS and COX-2, activation of NF-κB (A), expression of inflammatory cytokines (B), and production of PGE2 and NO (C) in CIA mice. Arthritis was induced by the first intradermal injection of bovine type II collagen and the second intradermal injection of collagen with complete Freund’s adjuvant at the base of the tail of DBA/1 J mice. Test agents (AC, 20 mg/kg DW2007; IB, 50 mg/kg ibuprofen; or CIA, vehicle alone) were orally administrated daily for 20 days from day 1 after the second injection. Proteins were determined in the paw joint tissues by immunoblotting. Cytokines were determined in the paw joint tissues by ELISA (IL-1β, IL-10, IL-17a, TNF-α, and PGE2). NO was measured using Griess reagent. All data indicate mean ± SD (n=7). # p <0.05 vs. control group. * p <0.05 vs. CIA group.

Article Snippet: The reaction mixture were filtered and T cells were isolated using a Pan T cell Isolation Kit II, fixed, stained with anti-Foxp3 or anti-IL-17A antibodies (Biogems, CA, USA) and analyzed by flow cytometry.

Techniques: Expressing, Activation Assay, Injection, Adjuvant, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Cell reports

Article Title: METTL14-dependent m 6 A modification controls iNKT cell development and function

doi: 10.1016/j.celrep.2022.111156

Figure Lengend Snippet: (A) Representative histogram of CD1d expression on DP thymocytes. CD1d was stained with α-CD1d or isotype control antibody (n = 8). (B) IL-2 detected by ELISA following 48-h co-culture of iNKT cell hybridoma DN32.D3 with irradiated thymocytes pulsed with α-GalCer ranging from 200 ng/mL to 12.5 ng/mL. Data representative of three independent experiments. (C) Flow cytometric analysis of iNKT cells in the Jα18 −/− recipient mice after 6 weeks of reconstitution with 1:1 mixture of bone marrow cells from WT (CD45.1) and T- Mettl14 −/− (CD45.2). (D) Quantification of iNKT cell reconstitution in thymus, spleen, and liver in bone marrow chimera recipients (n = 7). SEM is shown. ***p < 0.001.

Article Snippet: Purified rat anti-mouse IL-2 , BD Biosciences , RRID: AB_395383.

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Irradiation

Journal: Cell reports

Article Title: METTL14-dependent m 6 A modification controls iNKT cell development and function

doi: 10.1016/j.celrep.2022.111156

Figure Lengend Snippet: (A) Relative expression of Cish in naive and activated (anti-CD3/anti-CD4) DP thymocytes (n = 5–9). (B) Relative expression of Cish in the m 6 A RNA immunoprecipitation of total thymocytes RNA in WT mice by qPCR quantification (n = 4). (C) Intracellular calcium flux in DP thymocytes in response to crosslinking of anti-CD3/anti-CD4 in T- Mettl14 −/− mice. (D and E) Quantification of maximum calcium flux on crosslinking and addition of Ca 2+ in T- Mettl14 −/− DP cells (n = 5–6). (F) Representative staining of thymic iNKT cell expansion at day 3 after stimulation with IL-2 or IL-15. (G and H) Percentage and cell number of thymic iNKT cells on D0 and D3 after stimulation with IL-2 or IL-15 (n = 5). (I) Representative figure of cell trace distribution in thymic iNKT cells on day 3 post-stimulation with IL-2 or IL-15 (n = 5–6). Unstimulated thymocytes were used as controls. (J) Bar graph of pSTAT5 in iNKT thymocytes after 20 min of IL-15 stimulation (n = 5). SEM is shown. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Purified rat anti-mouse IL-2 , BD Biosciences , RRID: AB_395383.

Techniques: Expressing, Immunoprecipitation, Staining

Journal: Cell reports

Article Title: METTL14-dependent m 6 A modification controls iNKT cell development and function

doi: 10.1016/j.celrep.2022.111156

Figure Lengend Snippet: (A) Dot plots of IFN-γ and IL-4 in residual iNKT cells from T- Mettl14 −/− mice after in vivo α -GalCer stimulation. (B) Quantification of the percentage of IFN-γ and IL-4 producing iNKT cells in α-GalCer-immunized WT and T- Mettl14 −/− mice (n = 4). (C) Expression of METTL14 in DN32.D3 cells transduced with lentivirus coding Mettl14 -specific shRNA (sh Mettl14 -1 or sh Mettl14 -2) or control shRNA (shNC). (D) m 6 A level in mRNA of DN32.D3 cells transduced with sh Mettl14 -2 and shNC. (E) Relative expression of Mettl14 and Cish in DN32.D3-sh Mettl14 -2 in medium or stimulation with α-GalCer for 24 h. (F) Production of IL-2 in DN32.D3-sh Mettl14 -2 after stimulation with α-GalCer for 24 h quantified by ELISA. (G) Intracellular calcium flux in DN32.D3-sh Mettl14 -2 cells in response to α-GalCer/Ca 2+ . (H) Quantification of maximum calcium flux upon α-GalCer stimulation. Data representative of three to six independent experiments. shNC or sh Mettl14 -2-treated DN32.D3 were spin-transduced with retrovirus carrying sh Cish or shNC. Zsgreen + cells were sorted and cultured. (I) Relative expression of Cish in the indicated groups. (J) IL-2 production in shNC/shNC-treated (‘‘WT’’ control) and shNC and shCish-treated Mettl14 KD DN32.D3 cells after stimulation with α-GalCer for 24 h. Vα14 Tg splenocytes were nucleofected with rCas9/gRNA complex and maintained in complete RPMI supplemented with IL-2 for 3 days. (K) Mettl14 and Cish expression by qPCR on day 3 after nucleofection. (L) Absolute cell numbers of iNKT and CD4 + T cells after nucleofection. (M) IFN-γ production in iNKT and CD4 + T cells after nucleofection (n = 3–4). SEM is shown. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Purified rat anti-mouse IL-2 , BD Biosciences , RRID: AB_395383.

Techniques: In Vivo, Expressing, Transduction, shRNA, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cell reports

Article Title: METTL14-dependent m 6 A modification controls iNKT cell development and function

doi: 10.1016/j.celrep.2022.111156

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Purified rat anti-mouse IL-2 , BD Biosciences , RRID: AB_395383.

Techniques: Purification, shRNA, Recombinant, Enzyme-linked Immunosorbent Assay, Staining, SYBR Green Assay, Software, Chromogenic In Situ Hybridization

Journal:

Article Title: Transcriptome Analysis of Murine Macrophages in Response to Infection with Streptococcus pyogenes Reveals an Unusual Activation Program ▿

doi: 10.1128/IAI.00181-07

Figure Lengend Snippet: Genes differentially expressed in resident macrophages at 1 h postinfection with S. pyogenes

Article Snippet: In brief, 96-well microtiter plates were coated overnight at 4°C with purified rat anti-mouse anti-IL-6 capture antibody (Pharmingen, San Diego, CA) at 2 μg/ml in sodium bicarbonate buffer.

Techniques: Zinc-Fingers, Binding Assay

Journal:

Article Title: Transcriptome Analysis of Murine Macrophages in Response to Infection with Streptococcus pyogenes Reveals an Unusual Activation Program ▿

doi: 10.1128/IAI.00181-07

Figure Lengend Snippet: Confirmation of microarray data by RT-PCR and protein expression. (A) RT-PCR analysis of selected gene transcription in resident macrophages uninfected or infected with S. pyogenes. Uninfected samples were loaded in lanes 1, and infected samples were loaded in lanes 2. β-Actin expression served as a control. (B) IL-6 protein expression by S. pyogenes-infected macrophages. Resident macrophages were isolated from the peritoneal cavity of mice after 1 h of infection with S. pyogenes and cultured in vitro for 2 h. Noninfected macrophages were used as a control. The levels of IL-6 in the supernatants were determined by ELISA. Each column represents the mean ± standard deviation of triplicate samples obtained from three independent experiments. P < 0.0001 (ANOVA).

Article Snippet: In brief, 96-well microtiter plates were coated overnight at 4°C with purified rat anti-mouse anti-IL-6 capture antibody (Pharmingen, San Diego, CA) at 2 μg/ml in sodium bicarbonate buffer.

Techniques: Microarray, Reverse Transcription Polymerase Chain Reaction, Expressing, Infection, Isolation, Cell Culture, In Vitro, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Scientific Reports

Article Title: Response kinetics reveal novel features of ageing in murine T cells

doi: 10.1038/s41598-019-42120-1

Figure Lengend Snippet: Duration and strength of stimulation determine ageing-related cytokine profiles. Cytokine levels of IL-2 ( a ), IL-4 ( b ), IL-5 ( c ), IL-10 ( d ), IL-17 ( e ), IFN-γ ( f ), and TNF-α ( g ) were measured in supernatant of splenocytes exposed for two or four days to an intermediate (int) or high stimulatory strength. Data represent one experiment with young (n = 6, 2 months old) and aged (n = 6, 22–24 months old) mice. Mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 for difference between young and old mice at time points indicated using Two-way ANOVA.

Article Snippet: Cells were labelled intracellularly with the following antibodies according to the FoxP3 Transcription Factor staining buffer set protocol (eBioscience): anti-CD3zeta-FITC (clone H146-968) (Abcam, Cambridge, Cambridgeshire, UK); anti-CTLA-4-BV605 (clone UC10-4B9), and anti-TNF-α-BV785 (clone MP6-XT22) (BioLegend); anti-FoxP3-eFluor660 (clone 150D/E4), anti-GARP-PE (clone YGIC86), anti-IFN-γ-PE-Cy7 (clone XMG1.2), and IL-4-PE (clone 11B11) (eBioscience); anti-IL-5-PE (clone TRFK5) (BD Pharmingen).

Techniques: