il1rl1 alexa fluor 700  (R&D Systems)

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