monoclonal rat anti st2 il 33r  (R&D Systems)

Journal: bioRxiv

Article Title: Epithelial and stromal remodeling in murine Crohn’s disease-like ileitis

doi: 10.1101/2025.11.08.684908

Figure Lengend Snippet: (A) Epithelial-only UMAP colored by unsupervised clusters (total nuclei indicated; e.g., 47,103 after QC). (B) Dot plot of canonical epithelial markers validating lineage identities. (C) Sample-level composition of the epithelial compartment across AKR1–3 and SAMP1–4, summarized as proportions of absorptive, secretory, and ISC categories. (D) Epithelial UMAPs annotated with subclusters for AKR and SAMP (Enterocyte1–8, Goblet1–2, Paneth1–2, tuft, EEC, ISC/late-ISC. (E) Distribution of Pcsk6 expression across epithelial subclusters. (F) Functional enrichment of the top 100 marker genes for the Enterocyte7 subcluster using Metascape. (G) Il1rl1 expression in AKR and SAMP epithelium plotted separately for visualization of this differentially expressed gene (DEG) in UMAP space. Color represents normalized expression compared with all genes across all epithelial cells. (H) Immunohistochemistry for ST2 (encoded by Il1rl1 ) in AKR and SAMP ileum with inset magnifications; scale bars as labeled.

Article Snippet: Immunohistochemical (IHC) staining was performed using unconjugated polyclonal anti-mouse antibodies for DCLK1 (1:100, ab31704; Abcam, Cambridge, UK) or ST2 (AF1004; R&D Systems).

Techniques: Expressing, Functional Assay, Marker, Immunohistochemistry, Labeling

Journal: Communications Biology

Article Title: Macrophage-derived IL-1β directs fibroblast progenitor cell fate via metabolic reprogramming in wound healing

doi: 10.1038/s42003-025-08754-w

Figure Lengend Snippet: A UMAP visualizations display a distinct expression pattern of Il1rl1 , predominantly observed in C22 fibroblasts in oral mucosa and skin. B Violin plots reveal significantly elevated Il1rl1 expression levels in C22 fibroblasts. C Flow cytometric analysis confirms the notable expression of IL1RL1 in fibroblasts from oral mucosa and skin ( n = 4). Error bars represent SEM. D CytoTRACE analysis illustrates the stemness potential, highlighting C22 as potential fibroblast progenitors due to its highest stemness score. E , F Violin plots demonstrate an increased expression of Cd44 and Vcam1 ( Cd106 ) in Cluster 22 fibroblast subsets from both oral mucosa and skin. G Monocle 3 cell trajectory analysis, along with ( H ) RNA Velocity analysis, indicates that Cluster 22 fibroblasts are at the initial differentiation point, poised to mature into various fibroblast subsets.

Article Snippet: Antibodies were used for flowcytometry: PDGFRA-PE (135905, Biolegend, 1:100), Ki67- Alexa Fluor 700 (56-5698-82, Invitrogen, 1:100), CD45-FITC (103108, Biolegend, 0.5:100), IL-1β-PerCP 710 (46-7114-82, Invitrogen, 1:100), IL1RL1-Alexa Fluor 700 (FAB10041N, R&D systems, 1:100), PP65-APC (MA5-37167, Invitrogen, 1:100), PGK1 (Polyclonal Rabbit IgG, PA5-28612, Invitrogen, 0.5:100), NDUFA4 (Polyclonal Rabbit IgG, PA5-51021, Invitrogen, 1:100), FITC-conjugated Goat Anti-Rabbit IgG (SA00003-2, Proteintech, 1:300).

Techniques: Expressing

Journal: Communications Biology

Article Title: Macrophage-derived IL-1β directs fibroblast progenitor cell fate via metabolic reprogramming in wound healing

doi: 10.1038/s42003-025-08754-w

Figure Lengend Snippet: A Gene Set Enrichment Analysis (GSEA) plots highlight the activation of the oxidative phosphorylation pathway in Cluster 22 of oral mucosae (left), in contrast to ( B ) the glycolysis pathway predominating in skin fibroblasts (right). C Violin plots illustrate elevated expression levels of Ndufa4 in Cluster 22 ( Il1rl1 + fibroblasts) within the oral mucosa, and ( D ) increased Pgk1 expression in skin fibroblasts post-injury. E Flow cytometry confirms the upregulation of NDUFA4 in Il1rl1 + fibroblasts from oral mucosa, and ( F ) heightened PGK1 expression in Il1rl1 + fibroblasts from skin 48 h post-injury, with significance denoted as * p < 0.05, ** p < 0.01, and ‘ns’ indicating no significant difference ( n = 3). Error bars represent SEM.

Article Snippet: Antibodies were used for flowcytometry: PDGFRA-PE (135905, Biolegend, 1:100), Ki67- Alexa Fluor 700 (56-5698-82, Invitrogen, 1:100), CD45-FITC (103108, Biolegend, 0.5:100), IL-1β-PerCP 710 (46-7114-82, Invitrogen, 1:100), IL1RL1-Alexa Fluor 700 (FAB10041N, R&D systems, 1:100), PP65-APC (MA5-37167, Invitrogen, 1:100), PGK1 (Polyclonal Rabbit IgG, PA5-28612, Invitrogen, 0.5:100), NDUFA4 (Polyclonal Rabbit IgG, PA5-51021, Invitrogen, 1:100), FITC-conjugated Goat Anti-Rabbit IgG (SA00003-2, Proteintech, 1:300).

Techniques: Activation Assay, Phospho-proteomics, Expressing, Flow Cytometry

Journal: Communications Biology

Article Title: Macrophage-derived IL-1β directs fibroblast progenitor cell fate via metabolic reprogramming in wound healing

doi: 10.1038/s42003-025-08754-w

Figure Lengend Snippet: A GSEA enrichment plots illustrate the activation of the IL-1 signaling pathway, and ( B ) the IL-1 family signaling pathway in C22 fibroblasts from the oral mucosa post-injury, but not skin. C A violin plot reveals elevated Nfkb1 expression in C22 fibroblasts within the oral mucosa compared to the skin following injury. D Flow cytometric analysis confirms increased NFκB/pP65 expression in C22 fibroblasts of the oral mucosa post-injury, with *** p < 0.001 and ‘ns’ denoting no significant difference ( n = 4). Error bars represent SEM. E Representative immunofluorescent images showing the distribution of IL1RL1 + cells in oral muocsa and skin as well as the co-expression of IL1RL1 + and NFκB / PP65 in the injured oral mucosa. Scale bar, 50 μm.

Article Snippet: Antibodies were used for flowcytometry: PDGFRA-PE (135905, Biolegend, 1:100), Ki67- Alexa Fluor 700 (56-5698-82, Invitrogen, 1:100), CD45-FITC (103108, Biolegend, 0.5:100), IL-1β-PerCP 710 (46-7114-82, Invitrogen, 1:100), IL1RL1-Alexa Fluor 700 (FAB10041N, R&D systems, 1:100), PP65-APC (MA5-37167, Invitrogen, 1:100), PGK1 (Polyclonal Rabbit IgG, PA5-28612, Invitrogen, 0.5:100), NDUFA4 (Polyclonal Rabbit IgG, PA5-51021, Invitrogen, 1:100), FITC-conjugated Goat Anti-Rabbit IgG (SA00003-2, Proteintech, 1:300).

Techniques: Activation Assay, Expressing

Journal: Communications Biology

Article Title: Macrophage-derived IL-1β directs fibroblast progenitor cell fate via metabolic reprogramming in wound healing

doi: 10.1038/s42003-025-08754-w

Figure Lengend Snippet: A A dot plot demonstrates the enriched expression of Il1r1 in Cluster 22 relative to other fibroblast subsets. B CellChat analysis of ligand-receptor interactions within the IL-1 signaling network between Cluster 2 and fibroblast subsets, showing the most potent IL-1 signaling from C2 (macrophages) to C22 ( Il1rl1 + fibroblasts) in the oral mucosa. The color gradient reflects the interaction intensity. C Flow cytometric analysis confirms increased NFκB/PP65 expression in the fibroblasts of the oral mucosa with the same level of exogenous IL-1B, with * p < 0.001 and ‘ns’ denoting no significant difference ( n = 3). Error bars represent SEM. D Measurement of oxygen consumption rate (OCR) in oral mucosa and skin fibroblasts treated with exogenous IL-1β, respectively. Gray lines indicated the time points of adding oligomycin, carbonyl cyanite-4 (trifluoromethoxy) phenylhydrazone (FCCP), and Rotenone. E , F Statistical analysis of basal respiration ( E ) and ATP production ( F ), ** p < 0.01 and ‘ns’ indicating no significant difference ( n = 4). Error bars represent standard deviation (SD). G Measurement of extracellular acidification rate (ECAR) in oral mucosa and skin fibroblasts treated with exogenous IL-1β, respectively. Gray lines indicated the time points of adding glucose, oligomycin and 2-DG. H , I Statistical analysis of glycolytic capacity ( H ) and glycolysis ( I ), ** p < 0.01, ** p < 0.05, and ‘ns’ indicating no significant difference ( n = 5). Error bars represent SD. J Flow cytometric analysis confirms decreased NFκB/PP65 expression in C22 fibroblasts of the injured oral mucosa after applying the proteasome inhibitor, with * p < 0.05 and ‘ns’ denoting no significant difference ( n = 6). Error bars represent SEM. K Flow cytometric analysis confirms decreased NDUFA4 expression in C22 fibroblasts of the injured oral mucosa after applying the proteasome inhibitor, with * p < 0.05 and ‘ns’ denoting no significant difference ( n = 6). Error bars represent SEM. L Flow cytometric analysis confirms decreased PDGFRA level in the injured oral mucosa after applying the proteasome inhibitor, with *** p < 0.001 and ‘ns’ denoting no significant difference ( n = 6). Error bars represent SEM. M Histological analysis of oral mucosa after treatment with MG132 using H&E staining. Bar = 300 μm.

Article Snippet: Antibodies were used for flowcytometry: PDGFRA-PE (135905, Biolegend, 1:100), Ki67- Alexa Fluor 700 (56-5698-82, Invitrogen, 1:100), CD45-FITC (103108, Biolegend, 0.5:100), IL-1β-PerCP 710 (46-7114-82, Invitrogen, 1:100), IL1RL1-Alexa Fluor 700 (FAB10041N, R&D systems, 1:100), PP65-APC (MA5-37167, Invitrogen, 1:100), PGK1 (Polyclonal Rabbit IgG, PA5-28612, Invitrogen, 0.5:100), NDUFA4 (Polyclonal Rabbit IgG, PA5-51021, Invitrogen, 1:100), FITC-conjugated Goat Anti-Rabbit IgG (SA00003-2, Proteintech, 1:300).

Techniques: Expressing, Standard Deviation, Staining

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) A schematic of experimental design. CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 alone, IL2 and IL-33, or anti-CD3/CD28 under Th2 polarizing conditions for five days. (B) Representative flow plot of CD4 T cells immediately following isolation from secondary lymphoid organs of naïve or Hpoly -infected mice. (C) CD4 T cells from secondary lymphoid organs of Hpoly -infected mice were cultured under indicated conditions. Frequencies and numbers of GATA3+ST2+ cells were quantified. (D) Representative flow plot of CD4 T cells right after isolation from secondary lymphoid organs of Hpoly-infected 4get mice. (E) CD4 T cells from naïve or Hpoly-infected mice were cultured with IL2 and IL-33. (F) CD4 T cells from Hpoly-infected mice were cultured under the indicated conditions. Frequencies and numbers of GATA3+ST2+ cells were quantified after five days. (G) Abundance of amphiregulin in the culture supernatants from (F). (H) Splenocytes from naïve or Hpoly-infected mice cultured with IL2 alone or together with HES. Frequencies and numbers of GATA3+ cells was quantified. The data (B-H) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B-G) or two-way ANOVA with Sidak post hoc test. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Representative flow plots and quantification of ST2+GATA3 + in live CD45+TCRβ+CD4+FOXP3-T cells from mesenteric adipose tissue of Hpoly -infected mice or after five days of in vitro culture of CD4 T cells isolated from Hpoly-infected mice. Flow plots and quantification of ST2+IL4+(GFP) (B) or CD44+IL4+(GFP+) (C) cells cultured as above. (D) CD4 T cells from naïve or Hpoly -infected mice were cultured as in (A) and IL-33R+GATA3+ Th2 cells were quantified. (E) Representative flow plots and quantification of AREG+ cells within the IL4+(GFP+) population after 5hr restimulation with PMA/Iono in the presence of Brefeldin. The data (A-E) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (A-C, E) or two-way ANOVA with Sidak post hoc test (D).. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, *** p ≤ 0.0001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Infection, In Vitro, Isolation, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) CD44+IL4+ (GFP)+ early Th2 cells, CD44+IL4-(GPF-) activated cells, and naïve CD44-GFP-cells were FACS-isolated from 4get mice infected with Hpoly and cultured with IL2 and IL-33 for five days. Frequencies of GFP+ and GFP+ST2+ cells (B) as well as ST2 gMFI were quantified (C). The data (A-C) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B) or paired T-test (C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: RNAseq was performed on CD44+IL4+(GFP+) Th2 cells isolated from spleens and lymph nodes of Hpoly -infected mice or after five-day culture under indicated conditions. (A) Principal component analysis of RNAseq data. (B) Heatmap of hierarchical clustering of differentially expressed genes. (C) Top Hallmark pathways from GSEA for IL-33 v IL2 comparison. (D) Flow plots and quantification of pS6 expression in CD44+IL4(GFP+) Th2 cells. (E) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 in the presence of rapamycin. Frequencies and numbers of GATA3+ST2+ Th2 cells were quantified. (F) Levels of indicated cytokines after overnight culture in fresh media containing rapamycin. The data (D, E, F) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (D, E) or paired T-test (F). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Comparison, Expressing, Cell Culture

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Top Hallmark pathways from GSEA for IL-33 v IL2 comparison. (B) CD4 T cells were isolated from Hpoly -infected mice and cultured anti-CD3/CD28 in the presence of rapamycin. The mean fluorescence intensity of GATA3 and cell numbers were quantified. (C) Levels of indicated cytokines after overnight culture of anti-CD3/CD28-stimulated cells in fresh media containing rapamycin. (D) Quantification of results from SCENITH assay in GATA3+ Th2 cells. (E) Representative flow plot showing the expression of TSLPR and ST2 after five days of in vitro culture of CD4 T cells were isolated from Hpoly -infected mice. (F) Representative flow plot and quantification of mitotracker green and mitotracker deep red staining after five days of culture. The data (B-F) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B, E, F), or paired T-test (C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Comparison, Isolation, Infection, Cell Culture, Fluorescence, Expressing, In Vitro, Staining

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Heatmap representing the abundance of the top 100 detected metabolites. (B) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 for five days. The intracellular abundance of indicated amino acids in naïve and CD4+CD44+ activated cells was determined via LC/MS. (C) Heatmap depicting the expression of arginine transporters genes related in RNAseq data from . (D) CD4 T cells were isolated from Hpoly -infected mice and cultured with IL2 and IL-33 for five days in amino acid drop-out media. Frequencies and numbers of GATA3+ST2+ Th2 cells were quantified. (E) Representative histograms and quantification of CTV dilution of CD4 T cells cultured as in D. (F) After five days of culture with IL2 and IL-33 in control or arginine-depleted media, cells were restimulated with PMA/Iono and amphiregulin was quantified. (G) Representative histograms and quantification of ps6 levels in pS6-positive cells at day 1 of culture. The data (D-G) are representative of at least two independent experiments. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (D, E), paired T-test (F), or two-way ANOVA with Sidak post hoc test. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture, Liquid Chromatography with Mass Spectroscopy, Expressing, Control

Journal: bioRxiv

Article Title: IL-33 promotes transcriptional and metabolic adaptations of tissue-resident Th2 cells

doi: 10.1101/2025.07.09.663905

Figure Lengend Snippet: (A) Expresssion of Th2 cell related genes in scRNAseq dataset of T cells isolated from mAT of Hpoly -infected mice identifying cluster of GATA3+ST2+ Th2 cells (circled). CD4 T cells from Hpoly-infected mice were stimulated with anti-CD3/CD28 and treated with DFMO (B) or cultured in arginine-depleted media (C). Mean fluorescence intestity of GATA3 and cells numbers were quantified. The data (B,C) are representative of at least two independent experiments. Symbols in the quantified data represent independent biological replicates. Data were analyzed by one-way ANOVA with Tukey’s post hoc test (B, C). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Additionally, biotinylated antibodies were used for the staining ST2 (IL-33R) (MDB, clone DJ8) and amphiregulin (R&D Systems, #BAF989).

Techniques: Isolation, Infection, Cell Culture, Fluorescence