recombinant mouse il 1β  (MedChemExpress)

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

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

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

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

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

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A) Schematic of the primary mouse astrocyte cultures. (B) Representative immunofluorescence images of cultured astrocytes stained for GFAP, showing characteristic astrocyte morphology. (C) Overview of RNA-seq and capped small (cs)RNA-seq data generation from astrocytes. The schematic shows the typical distribution of csRNA-seq and RNA-seq at various genomic locations, which allows the identification of Transcriptional Start Site (TSSs) using HOMER2. (D) Volcano plot of RNA-seq differential expression in astrocytes treated with vehicle (Veh) or IL-1B (10 ng/mL, 1 h). (E) Pathway enrichment analysis of IL-1B-induced differentially expressed genes. (F) RT-qPCR validation of selected IL-1B-responsive genes in astrocytes

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: Immunofluorescence, Cell Culture, Staining, RNA Sequencing, Quantitative Proteomics, Quantitative RT-PCR, Biomarker Discovery

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: Genome browser views showing RNA-seq signal at the Cxcl1, Ccl20, Csf2 , and Il6 loci in astrocytes treated with vehicle or IL-1B. These representative examples illustrate the robust transcriptional induction of canonical inflammatory genes following IL-1B stimulation. Gene models and genomic coordinates are shown in mm10 .

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A) Pairwise correlation analysis of csRNA-seq replicates from untreated and IL-1B-treated astrocytes. (B) Distribution of strand-specific csRNA-seq reads around GENCODE-annotated TSSs, showing strong enrichment at annotated transcriptional start sites. (C) Genomic annotation of astrocytes TSRs, partitioned across promoter, intronic, intergenic, and other genomic features. (D) Average chromatin profiles at promoter-distal TSRs, showing ATAC-seq and H3K27ac enrichment around transcribed regulatory elements. (E) Average csRNA-seq signal centered on promoter-distal ATAC-seq peaks, comparing transcribed and non-transcribed accessible regions (No Tx: 225,150 peaks, Tx: 14,836 peaks). (F) Average mCH profiles at promoter-proximal TSRs, promoter-distal transcribed TSRs, and non-transcribed distal accessible regions in astrocytes (data from ). (G) Distribution of NFIA and TEAD4 ChIP-seq peaks found overlapping TSRs and ATAC-seq peaks in astrocytes. (H)ChIP-seq read density for NFIA and TEAD4 centered on csRNA-seq defined TSRs, showing transcription factor binding immediately upstream of the primary TSS.

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: ChIP-sequencing, Binding Assay

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A)Volcano plot of differentially regulated csRNA-seq signal at astrocyte TSRs following IL-1B stimulation, identifying induced and repressed regulatory elements. (B) Genome browser tracks depicting induction of promoter and distal regulatory enhancer transcription by csRNA-seq at the Ccl2 locus. (C) GREAT functional enrichment analysis of IL-1B induced TSRs. (D) Spatial clustering of IL-1B induced TSRs, depicting the density of TSRs in each category adjacent to Induced TSRs. (E) De novo motif enrichment analysis of IL-1B induced TSRs by HOMER. (F) Average csRNA-seq signal centered on promoter-distal NF-κB/p65 peaks, showing increased bidirectional transcription at p65-bound distal elements after IL-1B stimulation, consistent with eRNA induction. (G) Spatial density of NF-κB, AP1, and IRF motifs relative to TSRs (TF binding sites per bp per TSS), showing upstream enrichment of these motifs at IL-1B-induced TSRs compared with unchanged or repressed TSRs.

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: Functional Assay, Binding Assay

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A) Genome browser example of an IL-1B-induced locus (Cxcl10) in astrocytes, showing increased transcription initiation after stimulation. (B) Scatter plot comparing IL-1B-induced Log2 csRNA-seq changes at the promoter vs. RNA-seq changes across genes, highlighting genes regulated primarily at initiation (along x-axis) versus those showing stronger changes at the mRNA level (along y-axis). (C) Genome browser tracks at the Junb locus showing increased gene expression and RNAPII elongation in the gene body with limited change in promoter initiation and RNAPII promoter levels, consistent with regulation being mediated primarily at the level of transcription elongation rather than increased initiation. (D) RNAPII ChIP-seq levels at the promoters of IL-1B induced genes stratified by genes with minimal versus strong increases in csRNA-seq initiation activity. (E) Scatter plot comparing changes in overall TSR levels (Log2 Fold change, NT vs. IL-1B) versus their WIP score significance (the −Log10 p-value), identifying TSRs with altered initiation patterns independent of changes in total transcriptional output. (F) Representative examples of TSRs exhibiting a strong change in initiation pattern (top, WIP score −1.61, Lg10 p-value = 9.54e-05) versus a strong change in overall initiation levels with minimal change in initiation shape(WIP score −0.13, Log10 p-value = 0.84). (G) Scatter plot of TF motif enrichment in TSRs with significant changes in overall activity versus changes in TSS positions, highlighting differential associations of NF-κB, TEAD, and NFI motifs with these TSRs classes. (H) Venn diagram showing the overlap of ChIP-seq peaks for NFIA and TEAD4 before and after IL-1B stimulation in astrocytes. (I) Motif enrichment analysis of condition-specific (either Veh or IL-1B) TEAD4- and NFIA-bound regions, showing IL-1B-specific enrichment for inflammatory TF motifs, including NF-κB, IRF (IRF8), and AP1 (i.e. Fra1).

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: RNA Sequencing, Gene Expression, ChIP-sequencing, Activity Assay

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A) Venn diagram showing overlap between IL-1B-induced genes in astrocytes and KLA-induced genes in bone marrow-derived macrophages (BMDMs). (B) Venn diagram showing overlap between induced TSRs in astrocytes and BMDMs, revealing largely distinct stimulus-responsive enhancer landscapes despite partial overlap in induced genes. (C) Genome browser view of Tnfaip3 locus illustrating that astrocytes and BMDMs induce the same gene but use different enhancers upstream a shared promoter. (D) Top: Venn diagram showing overlap of NF-κB binding sites between activated astrocytes and BMDMs. Bottom: Fraction of astrocyte-specific, shared, and BMDM-specific p65 peaks associated with TSRs induced in astrocytes only, in both cell types, and in BMDM only. (E) Heatmap of de novo motif enrichment at cell type-specific induced TSRs, showing enrichment of lineage-associated motifs for each cell type. (F) Fraction of induced TSR classes bound by lineage-associated TFs, showing preferential association of astrocyte-induced TSRs with NFI and TEAD4 and macrophage-induced TSRs with PU.1 and CEBPα.

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: Derivative Assay, Binding Assay

Journal: bioRxiv

Article Title: Transcription initiation profiling defines the regulatory logic of astrocyte gene regulation

doi: 10.64898/2026.05.03.722406

Figure Lengend Snippet: (A) Average eRNA signal centered on astrocyte-specific, shared, and BMDM-specific NF-κBp65 peaks, showing cell type-matched induction of regulatory transcription at p65 bound sites. (B) Fraction of astrocyte-specific, shared, and BMDM-specific p65 peaks overlapping accessible chromatin regions in astrocytes or BMDMs, indicating that NF-κB recruitment occurs preferentially at cell-type-specific open chromatin regions. (C) Violin plots showing increase in ChIP-seq signal for p65, NFIA, TEAD4 at astrocyte IL-1B-induced TSRs and for p65, PU.1, and CEBPβ in macrophage KLA-induced TSRs.

Article Snippet: Two days before collection, the astrocytes were plated at a density of 500,000 cells per well on 4-well chamber slides to perform immunocytochemistry or a density of 5.5 million cells per 15 cm dish to be treated with 10ng/mL of recombinant human interleukin 1 beta (IL-1B) (Invivogen, Cat#rcyec-h) for 1hr before sample collection.

Techniques: ChIP-sequencing

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation changes the transcriptomic profile of BM-hMSCs. Schematic of experimental design A . BM-hMSCs were exposed to IL-1 β (3 replicates, 1 donor) or left unstimulated (3 replicates, 1 donor) for one hour, followed by bulk RNA sequencing. PCA plot was used to show the variance between the unstimulated control (green) and IL-1 β exposed (purple) samples B . Heatmap displaying the Z-score of the top 100 varying genes across all samples. Red color indicates higher expression of the genes and blue color indicates decreased expression C . Volcano plot of differentially expressed genes, showing their log2 fold change (X-axis) and -log10 adjusted p-values (Y-axis) D . IL-1 β Interleukin-1β, BM-hMSCs, Bone marrow derived human mesenchymal cells, PCA Principal component analysis, FC Fold change, NS not significant. Figure 1A was created using Biorender.com

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: RNA Sequencing, Control, Expressing, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation increases expression of neutrophil recruitment and NF-kB signaling genes by BM-hMSCs. Heatmap clustering of the Z-score of the top 20 differentially expressed genes between unstimulated control BM-hMSCs (3 replicates, 1 donor, green) and IL-1 β stimulated BM-hMSCs (3 replicates, 1 donor, purple). Red color indicates higher expression of the genes and blue color indicates decreased expression A . Protein expression of CXCL1 (10 replicates, 4 donors), CCL2 (4 replicates, 2 donors), CXCL5 (4 replicates, 2 donors), and CXCL8/IL-8 (7 replicates, 3 donors) in BM-hMSCs secretome measured by ELLA or ELISA. Data are presented as median, and statistical analysis was performed using unpaired t-test with Welch’s correction B . Bar plot of the top 20 significant Gene Ontology (GO) biological processes representing IL- 1β induced upregulated differentially expressed genes C IL-1 β, Interleukin-1β; BM-hMSCs, Bone marrow derived human mesenchymal cells; CCL2, chemokine (C–C motif) ligand 2; CXCL1, chemokine (C-X-C motif) ligand 1; CXCL2, chemokine (C-X-C motif) ligand 1; CXCL8, chemokine (C-X-C motif) ligand 8; ****, p < 0,0001

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulated BM-hMSCs increased neutrophil recruitment partly via the NF-kB signaling pathway. Schematic of experimental design of the neutrophil migration assay. Illustration was created with BioRender.com A . Number of neutrophils that migrated through the transwell membrane from the top well to the bottom towards control conditioned medium, conditioned medium from IL-1β stimulated BM-hMSCs, and conditioned medium from IL-1β with the NF-kB inhibitor stimulated BM-hMSCs. The experiment was performed using neutrophils from four different donors (each donor represent one data point) and the experiments were performed on three different days. B . Protein expression of CXCL1 was measured in conditioned medium from unstimulated BM-hMSCs (4 replicates, 1 donor, green), IL-1β stimulated BM-hMSCs (4 replicates, 1 donor, purple), and ILβ stimulated BM-hMSCs with 10 μM BAY 11–7082 (4 replicates, 1 donor, grey) C . Phospho-p65 levels were quantified and normalized to total p65. Data are expressed relative to control, which was set to 1 D . IL-1β Interleukin-1β BM-hMSCs Bone marrow-derived human mesenchymal cells, Ctrl control, CXCL1 chemokine (C-X-C motif) ligand 1, ns not significant; * p < 0,05; ****, p < 0,0001

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: Migration, Membrane, Control, Expressing, Derivative Assay