Review

recombinant human rh il 1β (R&D Systems)

R&D Systems is a verified supplier

R&D Systems manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

R&D Systems recombinant human rh il 1β

A. GM-CSF human macrophages were stimulated with LPS (50 ng/mL) for 18 h in glucose-, fructose- (10 mM), or sugar-free media. <t>IL-1β,</t> IL-6, and IL-23 levels were quantified by ELISA. For IL-1β detection, 4 mM ATP was added during the final 2 h (n = 6). B. IL1B mRNA expression was analyzed by RT-qPCR under identical conditions (n = 5). C. Immunoblotting of cell lysates and supernatants from (A) was performed to assess pro- and mature (m) forms of IL-1β and caspase-1 (Casp-1). Band intensities were quantified by densitometry and normalized to β-actin (n = 5). D–E. Glycolytic activity was assessed by Seahorse extracellular flux analysis of ECAR following sequential injection of glucose, fructose (10 mM), or no sugar, followed by oligomycin (2 µM) and 2-DG (50 mM). Data represents five technical replicates (n = 3). F. HIF-1α protein and mRNA levels were measured in LPS-stimulated GM-CSF macrophages under indicated sugar conditions (n = 3–4). G–J. SKG mice were injected with curdlan to induce arthritis and administered fructose in drinking water starting 3 days prior to immunization. Glucose levels in serum and synovial fluid (SF) were assessed at day 42 (n = 5–8). H–I. mRNA levels of Hif1a and HIF-1α target genes in synovial cells were analyzed by RT-qPCR (n = 5). J. Intracellular HIF-1α expression was measured by flow cytometry in CD45⁺CD11b⁺F4/80⁺ synovial cells (n = 9–10). Graphs represent mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by Mann–Whitney U test (A–C, E–J) or unpaired t-test (D, F).

Recombinant Human Rh Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 947 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/recombinant human rh il 1β/product/R&D Systems
Average 96 stars, based on 947 article reviews

recombinant human rh il 1β - by Bioz Stars, 2026-06

96/100 stars

Images

1) Product Images from "Fructose utilization by GM-CSF-differentiated macrophages aggravates autoimmune inflammation via MG-derived AGE–RAGE signaling"

Article Title: Fructose utilization by GM-CSF-differentiated macrophages aggravates autoimmune inflammation via MG-derived AGE–RAGE signaling

Journal: bioRxiv

doi: 10.64898/2026.01.26.701899

A. GM-CSF human macrophages were stimulated with LPS (50 ng/mL) for 18 h in glucose-, fructose- (10 mM), or sugar-free media. IL-1β, IL-6, and IL-23 levels were quantified by ELISA. For IL-1β detection, 4 mM ATP was added during the final 2 h (n = 6). B. IL1B mRNA expression was analyzed by RT-qPCR under identical conditions (n = 5). C. Immunoblotting of cell lysates and supernatants from (A) was performed to assess pro- and mature (m) forms of IL-1β and caspase-1 (Casp-1). Band intensities were quantified by densitometry and normalized to β-actin (n = 5). D–E. Glycolytic activity was assessed by Seahorse extracellular flux analysis of ECAR following sequential injection of glucose, fructose (10 mM), or no sugar, followed by oligomycin (2 µM) and 2-DG (50 mM). Data represents five technical replicates (n = 3). F. HIF-1α protein and mRNA levels were measured in LPS-stimulated GM-CSF macrophages under indicated sugar conditions (n = 3–4). G–J. SKG mice were injected with curdlan to induce arthritis and administered fructose in drinking water starting 3 days prior to immunization. Glucose levels in serum and synovial fluid (SF) were assessed at day 42 (n = 5–8). H–I. mRNA levels of Hif1a and HIF-1α target genes in synovial cells were analyzed by RT-qPCR (n = 5). J. Intracellular HIF-1α expression was measured by flow cytometry in CD45⁺CD11b⁺F4/80⁺ synovial cells (n = 9–10). Graphs represent mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by Mann–Whitney U test (A–C, E–J) or unpaired t-test (D, F).

Figure Legend Snippet: A. GM-CSF human macrophages were stimulated with LPS (50 ng/mL) for 18 h in glucose-, fructose- (10 mM), or sugar-free media. IL-1β, IL-6, and IL-23 levels were quantified by ELISA. For IL-1β detection, 4 mM ATP was added during the final 2 h (n = 6). B. IL1B mRNA expression was analyzed by RT-qPCR under identical conditions (n = 5). C. Immunoblotting of cell lysates and supernatants from (A) was performed to assess pro- and mature (m) forms of IL-1β and caspase-1 (Casp-1). Band intensities were quantified by densitometry and normalized to β-actin (n = 5). D–E. Glycolytic activity was assessed by Seahorse extracellular flux analysis of ECAR following sequential injection of glucose, fructose (10 mM), or no sugar, followed by oligomycin (2 µM) and 2-DG (50 mM). Data represents five technical replicates (n = 3). F. HIF-1α protein and mRNA levels were measured in LPS-stimulated GM-CSF macrophages under indicated sugar conditions (n = 3–4). G–J. SKG mice were injected with curdlan to induce arthritis and administered fructose in drinking water starting 3 days prior to immunization. Glucose levels in serum and synovial fluid (SF) were assessed at day 42 (n = 5–8). H–I. mRNA levels of Hif1a and HIF-1α target genes in synovial cells were analyzed by RT-qPCR (n = 5). J. Intracellular HIF-1α expression was measured by flow cytometry in CD45⁺CD11b⁺F4/80⁺ synovial cells (n = 9–10). Graphs represent mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by Mann–Whitney U test (A–C, E–J) or unpaired t-test (D, F).

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Injection, Flow Cytometry, MANN-WHITNEY

Image Search Results

Journal: bioRxiv

Article Title: Fructose utilization by GM-CSF-differentiated macrophages aggravates autoimmune inflammation via MG-derived AGE–RAGE signaling

doi: 10.64898/2026.01.26.701899

Figure Lengend Snippet: A. GM-CSF human macrophages were stimulated with LPS (50 ng/mL) for 18 h in glucose-, fructose- (10 mM), or sugar-free media. IL-1β, IL-6, and IL-23 levels were quantified by ELISA. For IL-1β detection, 4 mM ATP was added during the final 2 h (n = 6). B. IL1B mRNA expression was analyzed by RT-qPCR under identical conditions (n = 5). C. Immunoblotting of cell lysates and supernatants from (A) was performed to assess pro- and mature (m) forms of IL-1β and caspase-1 (Casp-1). Band intensities were quantified by densitometry and normalized to β-actin (n = 5). D–E. Glycolytic activity was assessed by Seahorse extracellular flux analysis of ECAR following sequential injection of glucose, fructose (10 mM), or no sugar, followed by oligomycin (2 µM) and 2-DG (50 mM). Data represents five technical replicates (n = 3). F. HIF-1α protein and mRNA levels were measured in LPS-stimulated GM-CSF macrophages under indicated sugar conditions (n = 3–4). G–J. SKG mice were injected with curdlan to induce arthritis and administered fructose in drinking water starting 3 days prior to immunization. Glucose levels in serum and synovial fluid (SF) were assessed at day 42 (n = 5–8). H–I. mRNA levels of Hif1a and HIF-1α target genes in synovial cells were analyzed by RT-qPCR (n = 5). J. Intracellular HIF-1α expression was measured by flow cytometry in CD45⁺CD11b⁺F4/80⁺ synovial cells (n = 9–10). Graphs represent mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by Mann–Whitney U test (A–C, E–J) or unpaired t-test (D, F).

Article Snippet: Cells were plated in 96-well U-bottom plates and stimulated with anti-CD3/CD28-coated beads for 7 days under the following cytokine conditions: recombinant human (rh) IL-1β (30 ng/mL), rhIL-6 (30 ng/mL), rhIL-23 (10 ng/mL), and rhTGF-β (10 ng/mL) (all from R&D Systems, Minneapolis, MN) for Th17 polarization, rhIL-2 (100 IU/mL; PeproTech) and rhIL-12 (10 ng/mL; R&D Systems) for Th1 polarization, and rhIL-2 (100 IU/mL) alone for non-polarizing control.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Injection, Flow Cytometry, MANN-WHITNEY

Following persistent stimulation of mesenchymal stem cells (MSCs) by tumor necrosis factor α (TNFα) + interleukin 1β (IL-1β), the resulting cells acquire elongated cancer-associated fibroblast (CAF)-like morphology. Human MSCs were exposed to TNFα (50 ng/mL) + IL-1β (0.5 ng/mL) or to vehicles. Cytokine concentrations were selected based on the considerations described in the Materials and Methods section. ( A ) At different time points, the cells were photographed by light microscopy. Images from a representative experiment out of n > 3 are presented. Bar, 50 μm. ( B ) Determination of cell characteristics using IN Cell technology in cells that were treated using the cytokines/vehicles for 18 days and were then subjected to IN Cell analysis. ( B1 ) Cell morphology was detected by calcein (green) and Hoechst (blue) staining. Images of cell morphology from a representative experiment out of n = 3 are presented. Bar, 100 µm. ( B2 ) Quantification of cell characteristics by the IN Cell technology. *** p < 0.001. The results of a representative experiment of n = 3 are presented.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of mesenchymal stem cells (MSCs) by tumor necrosis factor α (TNFα) + interleukin 1β (IL-1β), the resulting cells acquire elongated cancer-associated fibroblast (CAF)-like morphology. Human MSCs were exposed to TNFα (50 ng/mL) + IL-1β (0.5 ng/mL) or to vehicles. Cytokine concentrations were selected based on the considerations described in the Materials and Methods section. ( A ) At different time points, the cells were photographed by light microscopy. Images from a representative experiment out of n > 3 are presented. Bar, 50 μm. ( B ) Determination of cell characteristics using IN Cell technology in cells that were treated using the cytokines/vehicles for 18 days and were then subjected to IN Cell analysis. ( B1 ) Cell morphology was detected by calcein (green) and Hoechst (blue) staining. Images of cell morphology from a representative experiment out of n = 3 are presented. Bar, 100 µm. ( B2 ) Quantification of cell characteristics by the IN Cell technology. *** p < 0.001. The results of a representative experiment of n = 3 are presented.

Article Snippet: Based on preliminary titration studies performed in our lab and on other in vitro reports using TNFα [ , , ] and IL-1β [ , ], MSCs were stimulated by recombinant human (rh) TNFα (50 ng/mL; #300-01A, PeproTech, NJ, USA) and/or rhIL-1β (0.5 ng/mL; #200-01B, PeproTech).

Techniques: Light Microscopy, Cell Analysis, Staining

Persistent stimulation of MSCs by TNFα + IL-1β leads to prominent alterations in gene expression and in fibroblast-relevant transcriptional programs in the resulting CAF-like cells. Human MSCs were exposed to “persistent” stimulation by TNFα and/or IL-1β for 14 days or to “short” stimulation of 48 h (concentrations as in ) in three independent biological repeats, followed by RNAseq analyses. Control cells were treated using the vehicles of the cytokines. ( A ) Heat maps. Deseq-normalized counts, values were centered. ( B ) Venn diagrams. Upregulated genes: FC > 2, padj < 0.05; downregulated genes: FC < 0.5, padj < 0.05. ( C ) Fibroblast-relevant transcriptional programs that were significantly modified by persistent TNFα + IL-1β stimulation, at FC > 2 or FC < 0.5 and padj < 5 × 10 −10 , are demonstrated.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Persistent stimulation of MSCs by TNFα + IL-1β leads to prominent alterations in gene expression and in fibroblast-relevant transcriptional programs in the resulting CAF-like cells. Human MSCs were exposed to “persistent” stimulation by TNFα and/or IL-1β for 14 days or to “short” stimulation of 48 h (concentrations as in ) in three independent biological repeats, followed by RNAseq analyses. Control cells were treated using the vehicles of the cytokines. ( A ) Heat maps. Deseq-normalized counts, values were centered. ( B ) Venn diagrams. Upregulated genes: FC > 2, padj < 0.05; downregulated genes: FC < 0.5, padj < 0.05. ( C ) Fibroblast-relevant transcriptional programs that were significantly modified by persistent TNFα + IL-1β stimulation, at FC > 2 or FC < 0.5 and padj < 5 × 10 −10 , are demonstrated.

Techniques: Gene Expression, Control, Modification

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting CAF-like cells express typical CAF markers. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. The expressions of vimentin ( A ) and fibroblast activation protein (FAP) ( B ) were determined by confocal analyses. ( A1 , B1 ) Two representative images are demonstrated for each treatment, derived from a representative experiment out of n = 3. Bar, 25 μm. ( A2 , B2 ) Quantitative analyses of the fluorescence intensity performed by the ImageJ program on images of the representative experiment ( n ≥ 5 images for each treatment). ** p < 0.01.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting CAF-like cells express typical CAF markers. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. The expressions of vimentin ( A ) and fibroblast activation protein (FAP) ( B ) were determined by confocal analyses. ( A1 , B1 ) Two representative images are demonstrated for each treatment, derived from a representative experiment out of n = 3. Bar, 25 μm. ( A2 , B2 ) Quantitative analyses of the fluorescence intensity performed by the ImageJ program on images of the representative experiment ( n ≥ 5 images for each treatment). ** p < 0.01.

Techniques: Activation Assay, Derivative Assay, Fluorescence

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting CAF-like cells express typical CAF functions. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. The figure demonstrates the results of a collagen contraction assay, in which the change in collagen gel size was determined along the process. ( A ) Kinetics analyses of changes in collagen gel diameter compared to original gel size, measured by ImageJ. The average ± SEM of 5 independent experiments is demonstrated. *** p < 0.001. ( B ) An image from a representative experiment out of n = 5 is presented. Three wells are demonstrated for each treatment. The photo was taken at day 10 of the collagen contraction assay. Bar, 15.6 mm.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting CAF-like cells express typical CAF functions. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. The figure demonstrates the results of a collagen contraction assay, in which the change in collagen gel size was determined along the process. ( A ) Kinetics analyses of changes in collagen gel diameter compared to original gel size, measured by ImageJ. The average ± SEM of 5 independent experiments is demonstrated. *** p < 0.001. ( B ) An image from a representative experiment out of n = 5 is presented. Three wells are demonstrated for each treatment. The photo was taken at day 10 of the collagen contraction assay. Bar, 15.6 mm.

Techniques: Contraction Assay

Persistent stimulation with TNFα + IL-1β leads to the conversion of MSCs to inflammatory CAFs. ( A , B ) Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. ( A ) αSMA expression was determined by confocal analyses. ( A1 ) Two images of each treatment, derived from a representative experiment out of n > 3, are presented. Bar, 25 μm. ( A2 ) Quantitative analyses of the fluorescence intensity performed by the ImageJ program on images of the representative experiment ( n ≥ 5 images for each treatment). ** p < 0.01. ( B ) Cell proliferation was determined by cell counts at the end of the stimulation process. Average ± SD of n > 3 is presented. * p < 0.05. Time-dependent analysis of MSC cell numbers along the stimulation process are demonstrated in . ( C ) Expression of pro-inflammatory genes and secreted proteins, determined at the end of the stimulation process, is demonstrated. ( C1 ) mRNA expression was determined by transcriptome analyses performed at day 14, with three independent biological repeats. padj < 10 × 10 −10 –padj < 5 × 10 −271 , depending on the gene. ( C2 ) Expression of secreted proteins, determined by secretome analyses performed at days 18–19 with three independent biological repeats. p < 10 −3 – p < 7.5 × 10 −7 , depending on the protein. Data are presented as the fold induction of values obtained in cytokine-stimulated cells compared to vehicle-treated cells for all genes and proteins. demonstrate the 60 top upregulated or downregulated genes and proteins, obtained following persistent stimulation of the MSCs by TNFα + IL-1β.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Persistent stimulation with TNFα + IL-1β leads to the conversion of MSCs to inflammatory CAFs. ( A , B ) Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. ( A ) αSMA expression was determined by confocal analyses. ( A1 ) Two images of each treatment, derived from a representative experiment out of n > 3, are presented. Bar, 25 μm. ( A2 ) Quantitative analyses of the fluorescence intensity performed by the ImageJ program on images of the representative experiment ( n ≥ 5 images for each treatment). ** p < 0.01. ( B ) Cell proliferation was determined by cell counts at the end of the stimulation process. Average ± SD of n > 3 is presented. * p < 0.05. Time-dependent analysis of MSC cell numbers along the stimulation process are demonstrated in . ( C ) Expression of pro-inflammatory genes and secreted proteins, determined at the end of the stimulation process, is demonstrated. ( C1 ) mRNA expression was determined by transcriptome analyses performed at day 14, with three independent biological repeats. padj < 10 × 10 −10 –padj < 5 × 10 −271 , depending on the gene. ( C2 ) Expression of secreted proteins, determined by secretome analyses performed at days 18–19 with three independent biological repeats. p < 10 −3 – p < 7.5 × 10 −7 , depending on the protein. Data are presented as the fold induction of values obtained in cytokine-stimulated cells compared to vehicle-treated cells for all genes and proteins. demonstrate the 60 top upregulated or downregulated genes and proteins, obtained following persistent stimulation of the MSCs by TNFα + IL-1β.

Techniques: Expressing, Derivative Assay, Fluorescence

Following persistent stimulation of MSCs with TNFα + IL-1β, cancer-relevant programs are promoted in the resulting inflammatory CAFs. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles for 14 days in transcriptome analyses and for 18–19 days in secretome analyses. ( A ) Heat maps of protein expression determined by secretome analysis of three independent biological repeats, comparing cytokine-stimulated cells and control cells. ( B ) Disorder analysis of cancer-related genes and secreted proteins for which expression was modified in cytokine-stimulated cells compared to vehicle-treated cells. ( B1 ) Gene expression data were obtained by RNAseq analysis, in which genes were filtered using a cutoff of FC > 2 or FC < 0.5, padj < 5 × 10 −10 . ( B2 ) Data of secreted proteins were obtained by secretome analysis, in which proteins were filtered using a cutoff of FC ≥ 2 or FC ≤ 0.5, p < 0.05. The INGENUITY program was used to identify affected disorders. Indicated disorders obeyed the cutoff of p < 10 −17 in transcriptome analyses and p < 5 × 10 −8 in secretome analyses.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, cancer-relevant programs are promoted in the resulting inflammatory CAFs. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles for 14 days in transcriptome analyses and for 18–19 days in secretome analyses. ( A ) Heat maps of protein expression determined by secretome analysis of three independent biological repeats, comparing cytokine-stimulated cells and control cells. ( B ) Disorder analysis of cancer-related genes and secreted proteins for which expression was modified in cytokine-stimulated cells compared to vehicle-treated cells. ( B1 ) Gene expression data were obtained by RNAseq analysis, in which genes were filtered using a cutoff of FC > 2 or FC < 0.5, padj < 5 × 10 −10 . ( B2 ) Data of secreted proteins were obtained by secretome analysis, in which proteins were filtered using a cutoff of FC ≥ 2 or FC ≤ 0.5, p < 0.05. The INGENUITY program was used to identify affected disorders. Indicated disorders obeyed the cutoff of p < 10 −17 in transcriptome analyses and p < 5 × 10 −8 in secretome analyses.

Techniques: Expressing, Control, Modification, Gene Expression

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that induce morphological changes in luminal-A breast cancers (BC) cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. At the end of the stimulation period, cytokines were replaced by fresh cytokine-devoid media, and 48 h later, conditioned media (CM) were collected and administered to human luminal-A BC cells. ( A ) Tumor cell characteristics were quantified 4 days or 3 days after the addition of CM to MCF-7 cells ( A1 ) and to T47D cells, repectively ( A2 ) by IN Cell technology. The results of a representative experiment out of n = 3 are presented. *** p < 0.001. ( B ) Tumor cell morphology was photographed by confocal microscopy 2–3 days after the addition of CM, using phalloidin staining of actin filaments (green) and Hoechst staining of nuclei (blue). ( B1 ) MCF-7 cells. ( B2 ) T47D cells. Images from a representative experiment out of n = 3 are presented. Bar, 75 μm.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that induce morphological changes in luminal-A breast cancers (BC) cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. At the end of the stimulation period, cytokines were replaced by fresh cytokine-devoid media, and 48 h later, conditioned media (CM) were collected and administered to human luminal-A BC cells. ( A ) Tumor cell characteristics were quantified 4 days or 3 days after the addition of CM to MCF-7 cells ( A1 ) and to T47D cells, repectively ( A2 ) by IN Cell technology. The results of a representative experiment out of n = 3 are presented. *** p < 0.001. ( B ) Tumor cell morphology was photographed by confocal microscopy 2–3 days after the addition of CM, using phalloidin staining of actin filaments (green) and Hoechst staining of nuclei (blue). ( B1 ) MCF-7 cells. ( B2 ) T47D cells. Images from a representative experiment out of n = 3 are presented. Bar, 75 μm.

Techniques: Confocal Microscopy, Staining

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that promote scattering of luminal-A BC cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to mCherry-expressing MCF-7 ( A ) and T47D ( B ) human luminal-A BC cells, followed by determination of tumor cell scattering out of spheroids (MCF-7 cells) or the ability to form spheroids (T47D cells). ( A1 , B1 ) Representative images of spheroid formation at days 8 or 4 for MCF-7 cells and T47D cells are demonstrated, respectively. Bar, 200 µm. The results of a representative experiment out of n = 3 are presented. ( A2 , B2 ) Quantitative analyses of spheroid sizes performed by the ImageJ program on images of the representative experiment ( n ≥ 7 images for each treatment). *** p < 0.001.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that promote scattering of luminal-A BC cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to mCherry-expressing MCF-7 ( A ) and T47D ( B ) human luminal-A BC cells, followed by determination of tumor cell scattering out of spheroids (MCF-7 cells) or the ability to form spheroids (T47D cells). ( A1 , B1 ) Representative images of spheroid formation at days 8 or 4 for MCF-7 cells and T47D cells are demonstrated, respectively. Bar, 200 µm. The results of a representative experiment out of n = 3 are presented. ( A2 , B2 ) Quantitative analyses of spheroid sizes performed by the ImageJ program on images of the representative experiment ( n ≥ 7 images for each treatment). *** p < 0.001.

Techniques: Expressing

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that promote migration of luminal-A BC cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to mCherry-expressing MCF-7 and T47D human luminal-A BC cells. ( A ) Tumor cell migration was determined in fibronectin-coated transwells in response to serum-containing medium. ( A1 ) MCF-7 cells. ( A2 ) T47D cells. Images and quantitative analyses of a representative experiment out of n = 3, for each cell type, are presented. *** p < 0.001. ( B ) MCF-7 wound closure assay, performed in IncuCyte ® . ( B1 ) Representative images taken at 24 h. Bar, 150 µm. ( B2 ) The kinetics graphs of tumor cell migration demonstrate the proportion (%) of the original wound area that became covered by migrating cells at each time point. The results presented are mean ± SEM of 5 replicates for each treatment and are of a representative experiment out of n > 3. *** p < 0.001.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that promote migration of luminal-A BC cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to mCherry-expressing MCF-7 and T47D human luminal-A BC cells. ( A ) Tumor cell migration was determined in fibronectin-coated transwells in response to serum-containing medium. ( A1 ) MCF-7 cells. ( A2 ) T47D cells. Images and quantitative analyses of a representative experiment out of n = 3, for each cell type, are presented. *** p < 0.001. ( B ) MCF-7 wound closure assay, performed in IncuCyte ® . ( B1 ) Representative images taken at 24 h. Bar, 150 µm. ( B2 ) The kinetics graphs of tumor cell migration demonstrate the proportion (%) of the original wound area that became covered by migrating cells at each time point. The results presented are mean ± SEM of 5 replicates for each treatment and are of a representative experiment out of n > 3. *** p < 0.001.

Techniques: Migration, Expressing, Wound Closure Assay

Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between Ras-activating receptors and G protein-coupled receptors (GPCR) that signal via Gαi. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of ( A ) FTS, an inhibitor of Ras; ( B ) PTx, an inhibitor of Gαi; or ( C ) both inhibitors together. Control cells were treated by the vehicles of inhibitors. Inhibitor concentrations were selected based on the considerations described in the Materials and Methods section. Cell numbers and viability were not affected by the inhibitors. Then, kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in B. The results are presented as mean ± SEM of 8–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between Ras-activating receptors and G protein-coupled receptors (GPCR) that signal via Gαi. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of ( A ) FTS, an inhibitor of Ras; ( B ) PTx, an inhibitor of Gαi; or ( C ) both inhibitors together. Control cells were treated by the vehicles of inhibitors. Inhibitor concentrations were selected based on the considerations described in the Materials and Methods section. Cell numbers and viability were not affected by the inhibitors. Then, kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in B. The results are presented as mean ± SEM of 8–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented.

Techniques: Migration, Derivative Assay, Control

Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between the chemokine receptors CCR2, CCR5, and CXCR1/2. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of inhibitors. ( A ) Chemokine receptor inhibitors. ( A1 ) CCR2i = the CCR2 inhibitor CAS 445479–97-0; ( A2 ) CCR5i = the CCR5 inhibitor Maraviroc; and ( A3 ) CXCR1/2i = the CXCR1/2 inhibitor Reparixin. ( B ) Combined inhibitory measures directed to chemokine receptors compared to inhibition of Gαi by PTx performed in the same experiment. ( B1 ) All three chemokine receptor inhibitors together. ( B2 ) Inhibition by PTx. In all treatments, cell numbers and viability were not affected by the inhibitors. Kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in . The results are presented as mean ± SEM of 6–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by the CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented (with the exception of n = 2/3 in (A1)).

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between the chemokine receptors CCR2, CCR5, and CXCR1/2. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of inhibitors. ( A ) Chemokine receptor inhibitors. ( A1 ) CCR2i = the CCR2 inhibitor CAS 445479–97-0; ( A2 ) CCR5i = the CCR5 inhibitor Maraviroc; and ( A3 ) CXCR1/2i = the CXCR1/2 inhibitor Reparixin. ( B ) Combined inhibitory measures directed to chemokine receptors compared to inhibition of Gαi by PTx performed in the same experiment. ( B1 ) All three chemokine receptor inhibitors together. ( B2 ) Inhibition by PTx. In all treatments, cell numbers and viability were not affected by the inhibitors. Kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in . The results are presented as mean ± SEM of 6–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by the CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented (with the exception of n = 2/3 in (A1)).

Techniques: Migration, Derivative Assay, Inhibition, Control

Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between Ras-activating receptors and chemokine receptors. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of inhibitors. In the same experiment, different inhibitory conditions were used: ( A ) combined inhibition of CCR2, CCR5, and CXCR1/2 together, as in . ( B ) Inhibition of Ras-mediated signaling by FTS. ( C ) Combined inhibitory measures directed to chemokine receptors and Ras-mediated signaling. In all treatments, cell numbers and viability were not affected by the inhibitors (except for the inhibitory treatment included in part C, where 15–25% decrease in cell numbers was noted). Kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in . The results are presented as mean ± SEM of 8–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by the CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented.

Journal: Cancers

Article Title: Persistent Inflammatory Stimulation Drives the Conversion of MSCs to Inflammatory CAFs That Promote Pro-Metastatic Characteristics in Breast Cancer Cells

doi: 10.3390/cancers13061472

Figure Lengend Snippet: Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between Ras-activating receptors and chemokine receptors. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in ) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in and were administered to MCF-7 cells in the presence of inhibitors. In the same experiment, different inhibitory conditions were used: ( A ) combined inhibition of CCR2, CCR5, and CXCR1/2 together, as in . ( B ) Inhibition of Ras-mediated signaling by FTS. ( C ) Combined inhibitory measures directed to chemokine receptors and Ras-mediated signaling. In all treatments, cell numbers and viability were not affected by the inhibitors (except for the inhibitory treatment included in part C, where 15–25% decrease in cell numbers was noted). Kinetics analyses of wound closure assays were performed in IncuCyte ® , as described in . The results are presented as mean ± SEM of 8–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by the CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented.

Techniques: Migration, Derivative Assay, Inhibition, Control

(A) mRNA expression of IL-1β and IL-1RA was analyzed by the UALCAN tool. (B) Co-expression between IL-1β and IL-1RA analyzed by the GEPIA tool. (C) Protein expression of IL-1β and IL-1RA confirmed by IHC. (D) Protein expression of IL-1β and IL-1RA confirmed by western blotting. (E) Association etween IL-1β and OS/RFS analyzed by the GEPIA tool. IL, interleukin; OS, overall survival; RFS, recurrence-free survival.

Journal: Oncology Reports

Article Title: Interleukin 1β/1RA axis in colorectal cancer regulates tumor invasion, proliferation and apoptosis via autophagy

doi: 10.3892/or.2020.7475

Figure Lengend Snippet: (A) mRNA expression of IL-1β and IL-1RA was analyzed by the UALCAN tool. (B) Co-expression between IL-1β and IL-1RA analyzed by the GEPIA tool. (C) Protein expression of IL-1β and IL-1RA confirmed by IHC. (D) Protein expression of IL-1β and IL-1RA confirmed by western blotting. (E) Association etween IL-1β and OS/RFS analyzed by the GEPIA tool. IL, interleukin; OS, overall survival; RFS, recurrence-free survival.

Article Snippet: Recombinant human (rh) IL-1β protein and IL-1RA protein were purchased from R&D Systems, Inc.

Techniques: Expressing, Western Blot

(A) EMT-associated markers detected by western blotting and RT-qPCR. (B) Autophagy-associated markers detected by western blotting and RT-qPCR. (C) EMT-associated markers detected by western blotting and RT-qPCR after treatment of RAPA in the rhIL-1β group. *P<0.05, **P<0.01 and ***P<0.001. EMT, epithelial-mesenchymal transition; RT-qPCR, reverse transcription quantitative PCR; RAPA, rapamycin rh, recombinant human, IL, interleukin.

Journal: Oncology Reports

Article Title: Interleukin 1β/1RA axis in colorectal cancer regulates tumor invasion, proliferation and apoptosis via autophagy

doi: 10.3892/or.2020.7475

Figure Lengend Snippet: (A) EMT-associated markers detected by western blotting and RT-qPCR. (B) Autophagy-associated markers detected by western blotting and RT-qPCR. (C) EMT-associated markers detected by western blotting and RT-qPCR after treatment of RAPA in the rhIL-1β group. *P<0.05, **P<0.01 and ***P<0.001. EMT, epithelial-mesenchymal transition; RT-qPCR, reverse transcription quantitative PCR; RAPA, rapamycin rh, recombinant human, IL, interleukin.

Article Snippet: Recombinant human (rh) IL-1β protein and IL-1RA protein were purchased from R&D Systems, Inc.

Techniques: Western Blot, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, Recombinant

(A) Images of the subcutaneous xenografts. (B) Growth curve of xenografts in rhIL-1β and control groups. (C) Weight of the rhIL-1β and control groups before and after treatment. **P<0.01, ***P<0.001. rh, recombinant human; IL, interleukin.

Journal: Oncology Reports

Article Title: Interleukin 1β/1RA axis in colorectal cancer regulates tumor invasion, proliferation and apoptosis via autophagy

doi: 10.3892/or.2020.7475

Figure Lengend Snippet: (A) Images of the subcutaneous xenografts. (B) Growth curve of xenografts in rhIL-1β and control groups. (C) Weight of the rhIL-1β and control groups before and after treatment. **P<0.01, ***P<0.001. rh, recombinant human; IL, interleukin.

Article Snippet: Recombinant human (rh) IL-1β protein and IL-1RA protein were purchased from R&D Systems, Inc.

Techniques: Control, Recombinant

Review

Structured Review

Images

1) Product Images from "Fructose utilization by GM-CSF-differentiated macrophages aggravates autoimmune inflammation via MG-derived AGE–RAGE signaling"

Similar Products

Image Search Results