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recombinant mouse il 1β  (MedChemExpress)


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    Structured Review

    MedChemExpress recombinant mouse il 1β
    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
    Recombinant Mouse Il 1β, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 99/100, based on 2381 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+mouse+il+1%CE%B2/Ferrostatin-1/pmc13298421-109-4-11
    Average 99 stars, based on 2381 article reviews
    recombinant mouse il 1β - by Bioz Stars, 2026-09
    99/100 stars

    Images

    1) Product Images from "IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion"

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    Journal: Current Issues in Molecular Biology

    doi: 10.3390/cimb48060606

    Intercellular communication analysis predicts IL-1β as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
    Figure Legend Snippet: Intercellular communication analysis predicts IL-1β as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.

    Techniques Used: Expressing

    IL-1β induces ferroptotic stress and impairs SSPC function in vitro. ( A ) Schematic overview of SSPC isolation by fluorescence-activated cell sorting and subsequent in vitro validation experiments. ( B ) Flow cytometry gating strategy for isolating Lin − /DPP4 + SSPCs from mouse callus tissue at 7 days post-fracture. ( C ) Representative Western blot images showing the expression of ACSL4, COX2, 4-HNE, NF-κB p65, and EPAS1 in SSPCs treated with vehicle, erastin, or IL-1β. β-actin was used as the loading control. ( D ) Quantification of Western blot protein expression levels shown in panel C ( n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Oxidized lipids are indicated by increased green fluorescence and reduced red fluorescence. Scale bar = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs. Scale bar = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining and the relative FerroOrange fluorescence intensity ( n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of colony-forming unit efficiency and cell viability measured by CCK-8 assay ( n = 3). ( I ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression ( n = 3). ( J ) Representative images of tri-lineage differentiation assays, including Alizarin Red S staining for osteogenesis, Alcian Blue staining for chondrogenesis, and Oil Red O staining for adipogenesis. Scale bar = 200 μm. ( K ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression, including Runx2, Opn, Acan, Col2a1, Pparg, and Cebpa ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
    Figure Legend Snippet: IL-1β induces ferroptotic stress and impairs SSPC function in vitro. ( A ) Schematic overview of SSPC isolation by fluorescence-activated cell sorting and subsequent in vitro validation experiments. ( B ) Flow cytometry gating strategy for isolating Lin − /DPP4 + SSPCs from mouse callus tissue at 7 days post-fracture. ( C ) Representative Western blot images showing the expression of ACSL4, COX2, 4-HNE, NF-κB p65, and EPAS1 in SSPCs treated with vehicle, erastin, or IL-1β. β-actin was used as the loading control. ( D ) Quantification of Western blot protein expression levels shown in panel C ( n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Oxidized lipids are indicated by increased green fluorescence and reduced red fluorescence. Scale bar = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs. Scale bar = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining and the relative FerroOrange fluorescence intensity ( n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of colony-forming unit efficiency and cell viability measured by CCK-8 assay ( n = 3). ( I ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression ( n = 3). ( J ) Representative images of tri-lineage differentiation assays, including Alizarin Red S staining for osteogenesis, Alcian Blue staining for chondrogenesis, and Oil Red O staining for adipogenesis. Scale bar = 200 μm. ( K ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression, including Runx2, Opn, Acan, Col2a1, Pparg, and Cebpa ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Techniques Used: In Vitro, Isolation, Fluorescence, FACS, Biomarker Discovery, Flow Cytometry, Western Blot, Expressing, Control, Staining, CCK-8 Assay, Quantitative RT-PCR, Marker, Gene Expression, Comparison

    Pharmacological EPAS1 inhibition attenuates IL-1β-induced ferroptotic stress and differentiation impairment in primary SSPCs. ( A ) Representative images of Alcian Blue and Alizarin Red S staining in primary SSPCs treated with vehicle control, IL-1β, IL-1β plus PT2385, or PT2385 alone. Scale bars = 200 μm. ( B ) Representative Western blot images showing ACSL4, 4-HNE, EPAS1, and NF-κB p65 protein levels in SSPCs under the indicated treatments. GAPDH was used as the loading control. ( C ) qRT-PCR analysis of differentiation-related marker genes, including the chondrogenic markers Sox9 and Acan and the osteogenic markers Runx2 and Opn (n = 3). ( D ) Quantification of Western blot protein expression levels shown in panel ( B ) (n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Red fluorescence indicates non-oxidized lipid signal, and green fluorescence indicates oxidized lipid signal. Scale bars = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs under the indicated treatments. Scale bars = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining (n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of relative FerroOrange fluorescence intensity (n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.
    Figure Legend Snippet: Pharmacological EPAS1 inhibition attenuates IL-1β-induced ferroptotic stress and differentiation impairment in primary SSPCs. ( A ) Representative images of Alcian Blue and Alizarin Red S staining in primary SSPCs treated with vehicle control, IL-1β, IL-1β plus PT2385, or PT2385 alone. Scale bars = 200 μm. ( B ) Representative Western blot images showing ACSL4, 4-HNE, EPAS1, and NF-κB p65 protein levels in SSPCs under the indicated treatments. GAPDH was used as the loading control. ( C ) qRT-PCR analysis of differentiation-related marker genes, including the chondrogenic markers Sox9 and Acan and the osteogenic markers Runx2 and Opn (n = 3). ( D ) Quantification of Western blot protein expression levels shown in panel ( B ) (n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Red fluorescence indicates non-oxidized lipid signal, and green fluorescence indicates oxidized lipid signal. Scale bars = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs under the indicated treatments. Scale bars = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining (n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of relative FerroOrange fluorescence intensity (n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Techniques Used: Inhibition, Staining, Control, Western Blot, Quantitative RT-PCR, Marker, Expressing, Fluorescence, Comparison

    EPAS1 inhibition attenuates IL-1β-impaired bone regeneration in vivo. ( A ) Representative micro-CT three-dimensional reconstructions and micro-CT sectional images of fractured mouse femurs from vehicle control, IL-1β, IL-1β plus PT2385, and PT2385 alone groups at 28 days post-fracture. Scale bars = 1 mm. ( B ) Quantitative micro-CT analysis of bone volume (BV) and bone volume fraction (BV/TV) in the fracture callus region. ( C ) Quantitative histomorphometric analysis of bone area and callus area among the four treatment groups. ( D ) Representative Safranin O/Fast Green staining images of the fracture callus region showing callus organization, cartilage matrix, and newly formed bone tissue under the indicated treatments. Scale bars = 500 μm. Data are presented as mean ± SD; n = 6 mice per group. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.
    Figure Legend Snippet: EPAS1 inhibition attenuates IL-1β-impaired bone regeneration in vivo. ( A ) Representative micro-CT three-dimensional reconstructions and micro-CT sectional images of fractured mouse femurs from vehicle control, IL-1β, IL-1β plus PT2385, and PT2385 alone groups at 28 days post-fracture. Scale bars = 1 mm. ( B ) Quantitative micro-CT analysis of bone volume (BV) and bone volume fraction (BV/TV) in the fracture callus region. ( C ) Quantitative histomorphometric analysis of bone area and callus area among the four treatment groups. ( D ) Representative Safranin O/Fast Green staining images of the fracture callus region showing callus organization, cartilage matrix, and newly formed bone tissue under the indicated treatments. Scale bars = 500 μm. Data are presented as mean ± SD; n = 6 mice per group. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Techniques Used: Inhibition, In Vivo, Micro-CT, Control, Staining, Comparison

    Mendelian randomization provides exploratory genetic support linking IL-1β and EPAS1 to human bone nonunion risk. ( A ) Forest plot summarizing inverse-variance weighted Mendelian randomization estimates for the tested exposures and bone nonunion risk. Odds ratios and 95% confidence intervals are shown. ( B ) Scatter plots showing the associations between SNP effects on EPAS1 expression or IL-1β levels and SNP effects on bone nonunion risk. Lines indicate estimates from different Mendelian randomization methods. ( C ) Single-SNP forest plots showing individual SNP estimates for EPAS1 and IL-1β. The combined inverse-variance weighted estimates are shown at the bottom. ( D ) Leave-one-out analyses for EPAS1 and IL-1β, performed by sequentially excluding one SNP at a time. ( E ) Funnel plots used to assess potential directional pleiotropy in the Mendelian randomization analyses for EPAS1 and IL-1β. Red colors, red lines, and numbers indicate representative annotations used to highlight key features discussed in the main text.
    Figure Legend Snippet: Mendelian randomization provides exploratory genetic support linking IL-1β and EPAS1 to human bone nonunion risk. ( A ) Forest plot summarizing inverse-variance weighted Mendelian randomization estimates for the tested exposures and bone nonunion risk. Odds ratios and 95% confidence intervals are shown. ( B ) Scatter plots showing the associations between SNP effects on EPAS1 expression or IL-1β levels and SNP effects on bone nonunion risk. Lines indicate estimates from different Mendelian randomization methods. ( C ) Single-SNP forest plots showing individual SNP estimates for EPAS1 and IL-1β. The combined inverse-variance weighted estimates are shown at the bottom. ( D ) Leave-one-out analyses for EPAS1 and IL-1β, performed by sequentially excluding one SNP at a time. ( E ) Funnel plots used to assess potential directional pleiotropy in the Mendelian randomization analyses for EPAS1 and IL-1β. Red colors, red lines, and numbers indicate representative annotations used to highlight key features discussed in the main text.

    Techniques Used: Expressing

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    Transfection:

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    Plasmid Preparation:

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    Incubation:

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    Derivative Assay:

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    Western Blot:

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    Staining:

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    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
    Recombinant Mouse Il 1β Il 1f2 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
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    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
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    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
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    Intercellular communication analysis <t>predicts</t> <t>IL-1β</t> as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.
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    Image Search Results


    Intercellular communication analysis predicts IL-1β as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.

    Journal: Current Issues in Molecular Biology

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    doi: 10.3390/cimb48060606

    Figure Lengend Snippet: Intercellular communication analysis predicts IL-1β as a candidate upstream signal potentially associated with neutrophils. ( A ) Chord diagram showing predicted ligand–target interactions between sender cell populations and receiver SSPCs in the NonU microenvironment. Immune cell populations contributed prominently to the inferred SSPC-directed signaling network. ( B ) Dot plot showing the top predicted ligands received by SSPCs and their expression patterns across potential sender cell populations. Dot size indicates the percentage of ligand-expressing cells, and color indicates average expression. ( C ) Heatmap showing log-fold changes in prioritized ligands in their corresponding sender populations when comparing NonU with Frac conditions. ( D ) Heatmap showing the predicted regulatory potential of prioritized ligands on SSPC target genes. Candidate transcription factors are indicated. ( E ) GO enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis-related terms are highlighted. ( F ) KEGG enrichment analysis of predicted IL-1β target genes in SSPCs. Ferroptosis and fatty acid metabolism pathways are highlighted. Red arrows and red-colored labels indicate representative ligands, cell populations, target genes, transcription factors, or enriched terms highlighted and discussed in the main text.

    Article Snippet: Cells were treated with recombinant mouse IL-1β (10 ng/mL), Ferrostatin-1 (Fer-1; MedChemExpress, Monmouth Junction, NJ, USA; #HY-100579; 1 μM), or their combination for 24 h. Cells were then collected for Western blot analysis of ferroptosis-related proteins and, where indicated, FerroOrange staining to assess intracellular ferrous iron accumulation.

    Techniques: Expressing

    IL-1β induces ferroptotic stress and impairs SSPC function in vitro. ( A ) Schematic overview of SSPC isolation by fluorescence-activated cell sorting and subsequent in vitro validation experiments. ( B ) Flow cytometry gating strategy for isolating Lin − /DPP4 + SSPCs from mouse callus tissue at 7 days post-fracture. ( C ) Representative Western blot images showing the expression of ACSL4, COX2, 4-HNE, NF-κB p65, and EPAS1 in SSPCs treated with vehicle, erastin, or IL-1β. β-actin was used as the loading control. ( D ) Quantification of Western blot protein expression levels shown in panel C ( n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Oxidized lipids are indicated by increased green fluorescence and reduced red fluorescence. Scale bar = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs. Scale bar = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining and the relative FerroOrange fluorescence intensity ( n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of colony-forming unit efficiency and cell viability measured by CCK-8 assay ( n = 3). ( I ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression ( n = 3). ( J ) Representative images of tri-lineage differentiation assays, including Alizarin Red S staining for osteogenesis, Alcian Blue staining for chondrogenesis, and Oil Red O staining for adipogenesis. Scale bar = 200 μm. ( K ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression, including Runx2, Opn, Acan, Col2a1, Pparg, and Cebpa ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Journal: Current Issues in Molecular Biology

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    doi: 10.3390/cimb48060606

    Figure Lengend Snippet: IL-1β induces ferroptotic stress and impairs SSPC function in vitro. ( A ) Schematic overview of SSPC isolation by fluorescence-activated cell sorting and subsequent in vitro validation experiments. ( B ) Flow cytometry gating strategy for isolating Lin − /DPP4 + SSPCs from mouse callus tissue at 7 days post-fracture. ( C ) Representative Western blot images showing the expression of ACSL4, COX2, 4-HNE, NF-κB p65, and EPAS1 in SSPCs treated with vehicle, erastin, or IL-1β. β-actin was used as the loading control. ( D ) Quantification of Western blot protein expression levels shown in panel C ( n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Oxidized lipids are indicated by increased green fluorescence and reduced red fluorescence. Scale bar = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs. Scale bar = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining and the relative FerroOrange fluorescence intensity ( n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of colony-forming unit efficiency and cell viability measured by CCK-8 assay ( n = 3). ( I ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression ( n = 3). ( J ) Representative images of tri-lineage differentiation assays, including Alizarin Red S staining for osteogenesis, Alcian Blue staining for chondrogenesis, and Oil Red O staining for adipogenesis. Scale bar = 200 μm. ( K ) qRT-PCR analysis of osteogenic, chondrogenic, and adipogenic marker gene expression, including Runx2, Opn, Acan, Col2a1, Pparg, and Cebpa ( n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Article Snippet: Cells were treated with recombinant mouse IL-1β (10 ng/mL), Ferrostatin-1 (Fer-1; MedChemExpress, Monmouth Junction, NJ, USA; #HY-100579; 1 μM), or their combination for 24 h. Cells were then collected for Western blot analysis of ferroptosis-related proteins and, where indicated, FerroOrange staining to assess intracellular ferrous iron accumulation.

    Techniques: In Vitro, Isolation, Fluorescence, FACS, Biomarker Discovery, Flow Cytometry, Western Blot, Expressing, Control, Staining, CCK-8 Assay, Quantitative RT-PCR, Marker, Gene Expression, Comparison

    Pharmacological EPAS1 inhibition attenuates IL-1β-induced ferroptotic stress and differentiation impairment in primary SSPCs. ( A ) Representative images of Alcian Blue and Alizarin Red S staining in primary SSPCs treated with vehicle control, IL-1β, IL-1β plus PT2385, or PT2385 alone. Scale bars = 200 μm. ( B ) Representative Western blot images showing ACSL4, 4-HNE, EPAS1, and NF-κB p65 protein levels in SSPCs under the indicated treatments. GAPDH was used as the loading control. ( C ) qRT-PCR analysis of differentiation-related marker genes, including the chondrogenic markers Sox9 and Acan and the osteogenic markers Runx2 and Opn (n = 3). ( D ) Quantification of Western blot protein expression levels shown in panel ( B ) (n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Red fluorescence indicates non-oxidized lipid signal, and green fluorescence indicates oxidized lipid signal. Scale bars = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs under the indicated treatments. Scale bars = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining (n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of relative FerroOrange fluorescence intensity (n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Journal: Current Issues in Molecular Biology

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    doi: 10.3390/cimb48060606

    Figure Lengend Snippet: Pharmacological EPAS1 inhibition attenuates IL-1β-induced ferroptotic stress and differentiation impairment in primary SSPCs. ( A ) Representative images of Alcian Blue and Alizarin Red S staining in primary SSPCs treated with vehicle control, IL-1β, IL-1β plus PT2385, or PT2385 alone. Scale bars = 200 μm. ( B ) Representative Western blot images showing ACSL4, 4-HNE, EPAS1, and NF-κB p65 protein levels in SSPCs under the indicated treatments. GAPDH was used as the loading control. ( C ) qRT-PCR analysis of differentiation-related marker genes, including the chondrogenic markers Sox9 and Acan and the osteogenic markers Runx2 and Opn (n = 3). ( D ) Quantification of Western blot protein expression levels shown in panel ( B ) (n = 3). ( E ) Representative BODIPY 581/591 C11 fluorescence images showing lipid peroxidation in SSPCs. Red fluorescence indicates non-oxidized lipid signal, and green fluorescence indicates oxidized lipid signal. Scale bars = 50 μm. ( F ) Representative FerroOrange fluorescence images showing intracellular Fe 2+ levels in SSPCs under the indicated treatments. Scale bars = 50 μm. ( G ) Quantification of the red/green fluorescence intensity ratio from BODIPY 581/591 C11 staining (n = 3). A lower red/green ratio indicates increased lipid peroxidation. ( H ) Quantification of relative FerroOrange fluorescence intensity (n = 3). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Article Snippet: Cells were treated with recombinant mouse IL-1β (10 ng/mL), Ferrostatin-1 (Fer-1; MedChemExpress, Monmouth Junction, NJ, USA; #HY-100579; 1 μM), or their combination for 24 h. Cells were then collected for Western blot analysis of ferroptosis-related proteins and, where indicated, FerroOrange staining to assess intracellular ferrous iron accumulation.

    Techniques: Inhibition, Staining, Control, Western Blot, Quantitative RT-PCR, Marker, Expressing, Fluorescence, Comparison

    EPAS1 inhibition attenuates IL-1β-impaired bone regeneration in vivo. ( A ) Representative micro-CT three-dimensional reconstructions and micro-CT sectional images of fractured mouse femurs from vehicle control, IL-1β, IL-1β plus PT2385, and PT2385 alone groups at 28 days post-fracture. Scale bars = 1 mm. ( B ) Quantitative micro-CT analysis of bone volume (BV) and bone volume fraction (BV/TV) in the fracture callus region. ( C ) Quantitative histomorphometric analysis of bone area and callus area among the four treatment groups. ( D ) Representative Safranin O/Fast Green staining images of the fracture callus region showing callus organization, cartilage matrix, and newly formed bone tissue under the indicated treatments. Scale bars = 500 μm. Data are presented as mean ± SD; n = 6 mice per group. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Journal: Current Issues in Molecular Biology

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    doi: 10.3390/cimb48060606

    Figure Lengend Snippet: EPAS1 inhibition attenuates IL-1β-impaired bone regeneration in vivo. ( A ) Representative micro-CT three-dimensional reconstructions and micro-CT sectional images of fractured mouse femurs from vehicle control, IL-1β, IL-1β plus PT2385, and PT2385 alone groups at 28 days post-fracture. Scale bars = 1 mm. ( B ) Quantitative micro-CT analysis of bone volume (BV) and bone volume fraction (BV/TV) in the fracture callus region. ( C ) Quantitative histomorphometric analysis of bone area and callus area among the four treatment groups. ( D ) Representative Safranin O/Fast Green staining images of the fracture callus region showing callus organization, cartilage matrix, and newly formed bone tissue under the indicated treatments. Scale bars = 500 μm. Data are presented as mean ± SD; n = 6 mice per group. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Article Snippet: Cells were treated with recombinant mouse IL-1β (10 ng/mL), Ferrostatin-1 (Fer-1; MedChemExpress, Monmouth Junction, NJ, USA; #HY-100579; 1 μM), or their combination for 24 h. Cells were then collected for Western blot analysis of ferroptosis-related proteins and, where indicated, FerroOrange staining to assess intracellular ferrous iron accumulation.

    Techniques: Inhibition, In Vivo, Micro-CT, Control, Staining, Comparison

    Mendelian randomization provides exploratory genetic support linking IL-1β and EPAS1 to human bone nonunion risk. ( A ) Forest plot summarizing inverse-variance weighted Mendelian randomization estimates for the tested exposures and bone nonunion risk. Odds ratios and 95% confidence intervals are shown. ( B ) Scatter plots showing the associations between SNP effects on EPAS1 expression or IL-1β levels and SNP effects on bone nonunion risk. Lines indicate estimates from different Mendelian randomization methods. ( C ) Single-SNP forest plots showing individual SNP estimates for EPAS1 and IL-1β. The combined inverse-variance weighted estimates are shown at the bottom. ( D ) Leave-one-out analyses for EPAS1 and IL-1β, performed by sequentially excluding one SNP at a time. ( E ) Funnel plots used to assess potential directional pleiotropy in the Mendelian randomization analyses for EPAS1 and IL-1β. Red colors, red lines, and numbers indicate representative annotations used to highlight key features discussed in the main text.

    Journal: Current Issues in Molecular Biology

    Article Title: IL-1β/EPAS1-Associated Ferroptotic Stress Impairs Skeletal Stem/Progenitor Cell Function in Inflammation-Associated Fracture Nonunion

    doi: 10.3390/cimb48060606

    Figure Lengend Snippet: Mendelian randomization provides exploratory genetic support linking IL-1β and EPAS1 to human bone nonunion risk. ( A ) Forest plot summarizing inverse-variance weighted Mendelian randomization estimates for the tested exposures and bone nonunion risk. Odds ratios and 95% confidence intervals are shown. ( B ) Scatter plots showing the associations between SNP effects on EPAS1 expression or IL-1β levels and SNP effects on bone nonunion risk. Lines indicate estimates from different Mendelian randomization methods. ( C ) Single-SNP forest plots showing individual SNP estimates for EPAS1 and IL-1β. The combined inverse-variance weighted estimates are shown at the bottom. ( D ) Leave-one-out analyses for EPAS1 and IL-1β, performed by sequentially excluding one SNP at a time. ( E ) Funnel plots used to assess potential directional pleiotropy in the Mendelian randomization analyses for EPAS1 and IL-1β. Red colors, red lines, and numbers indicate representative annotations used to highlight key features discussed in the main text.

    Article Snippet: Cells were treated with recombinant mouse IL-1β (10 ng/mL), Ferrostatin-1 (Fer-1; MedChemExpress, Monmouth Junction, NJ, USA; #HY-100579; 1 μM), or their combination for 24 h. Cells were then collected for Western blot analysis of ferroptosis-related proteins and, where indicated, FerroOrange staining to assess intracellular ferrous iron accumulation.

    Techniques: Expressing