il1b  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: Metformin Modulates Cell Oxidative Stress to Mitigate Corticosteroid-Induced Suppression of Osteogenesis in a 3D Model

doi: 10.2147/JIR.S498888

Figure Lengend Snippet: Impact of different concentrations of prednisolone on macrophage viability and inflammation properties. ( A ) Histogram shows macrophages’ viability on day 7, and ( B ) representative fluorescent image of cells. ( C ) ELISA results of cytokine expressions of IL1b, IL6, TNFα, IL10 and CCL18. (“ns” for “no significant difference”, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: ELISA kits for measuring CCL18 (DY394) and IL1b (DY201-05) were ordered from R&D Systems.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Metformin Modulates Cell Oxidative Stress to Mitigate Corticosteroid-Induced Suppression of Osteogenesis in a 3D Model

doi: 10.2147/JIR.S498888

Figure Lengend Snippet: Metformin regulates macrophage functions. ( A and B ) Comparison of ROS level and ATP production in macrophages. ( C ) ELISA results of cytokine expressions of IL1b, IL6, TNFα, IL10 and CCL18. (“ns” for “no significant difference”, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: ELISA kits for measuring CCL18 (DY394) and IL1b (DY201-05) were ordered from R&D Systems.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: QPCR normalized expression values. The normalized gene expression values for IL1B ( a ) show a statistically significant upregulation in testis (T) (**, p < 0.01) compared to prostate (P) and seminal vesicles (SV). No significant differences in IL1B levels were observed between P and SV ( p > 0.05). Regarding IL1R1 ( b ), normalized mRNA expression levels reveal significant differences (***, p < 0.001) between T, P, and SV. The bars above the histograms represent the standard error. below the graphs shows the results from the 2% agarose gel electrophoresis. In the gel images, the first lane (left) contains a 50 bp DNA ladder (LD), followed by lanes with the qPCR amplicons for P, T, and SV. The last lane (right) shows the qPCR negative control (CTRL). ( a ) shows the gel of IL1B (183 bp) and 18S (489 bp) qPCR amplicons from P, T, and SV, while ( b ) shows the gel for IL1R1 (137 bp) and 18S (489 bp) amplicons from the same samples.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Gene Expression, Agarose Gel Electrophoresis, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: Immunopositivity for IL1B and IL1R1 in some organs of the reproductive tract of the male rabbits. Both IL1B and its receptor were observed in the cytoplasm of germinal cells, inside the convoluted seminiferous cells ( a , b ). Positivity is particularly evident in the basal portion of the epithelium where spermatogonia and first-order spermatocytes are found (*). In the P, a weak positivity for IL1B was observed in some secreting epithelial cells ( c ), while the positivity for the receptor was evident in a high number of the secreting epithelial cells ( d ). All glandular epithelial cells in the SV were immunopositive for IL1B ( e ) and IL1R1 ( f ). The immunopositivity for both IL1B and IL1R1 was diffused in the cytoplasm of all the epithelial cells. Sections were counterstained with hematoxylin.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: IL1B WB. The figure shows the molecular weight marker (10 to 250 kDa) on the left, only the bands corresponding to the target IL1B weights are cut. Bands 1, 2, 3, 4, and 5, positioned between 30 and 35 kDa, confirm the presence of the IL1B protein in the seminal plasma of the five rabbits involved in the experiment.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Molecular Weight, Marker, Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: IL1B ELISA. The figure shows the IL1B levels (ng) for total protein (mg). There were no statistical differences in IL1B expression between experimental group (EG) 1 (control), EG 4 (NGF addition), EG 5 (NGF + TRKA inhibitor), and EG 6 (NGF + p75NTR inhibitor). The bar above the histograms represents the dataset standard error. Equivalent letters indicate not statistically significant differences ( p > 0.05).

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: NGF ELISA. NGF protein expression levels of EG 1 (control) versus EG 2 (addition of IL1B) or versus EG 3 (addition of IL1B plus IL1R1 antagonist) show statistically significant differences. The bar above the histograms represents the dataset standard error. Different letters (a,b,c) indicate statistically significant differences ( p < 0.05). Equivalent letters indicate not statistically significant differences ( p > 0.05).

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: PGF2α and PGE2 ELISA. When compared with EG 2 (addition of IL1B) or with EG 7 (addition of IL1B plus COX inhibitor), the PGF2α and PGE2 expression levels in EG 1 (control) show statistically significant differences ( p ˂ 0.001). EG 2, EG 3 (IL1B plus IL1R1 antagonist), and EG 7 when confronted with each other show statistically significant differences ( p ˂ 0.001). Different letters (a,b,c) indicate statistically significant differences ( p < 0.05). Equivalent letters indicate not statistically significant differences ( p > 0.05).

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: IL1B is associated with the inflammatory response and NGF production in the rabbit uterus (U). The binding of IL1B to the extracellular domain of the IL1R1 membrane receptor allows the recruitment of the IL1R3 co-receptor. This interaction forms the IL1R1/IL1R3 heterodimer via the intracellular toll–interleukin-1 receptor (TIR) domains of the two receptor polypeptide chains, thereby initiating signal transduction. The IL1R1/IL1R3 receptor complex then recruits the adaptor protein myeloid differentiation primary response gene 88 (MyD88), which activates interleukin-1 receptor-associated kinase 1 (IRAK1) and tumor necrosis factor receptor-associated factor 6 (TRAF-6). This activation cascade stimulates the nuclear factor-kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signaling pathways. Upon activation, NF-κB translocates to the nucleus where it acts as a transcription factor, upregulating the gene expression of pro-inflammatory cytokines (e.g., IL-1, IL-6, and TNFα) and COX2 which is essential for the synthesis of PGE2 and PGF2α starting from the arachidonic acid. Similarly, IL1B-mediated activation of p38 MAPK induces the transcription factor cyclic AMP response element-binding protein (CREB), which promotes the transcription of COX2, pro-inflammatory cytokine, and NGF gene, explaining how the inflammatory cytokine IL1B enhances NGF production in the rabbit U. In addition to IL1B, NGF, through its receptors, p75NTR and TRKA, activate NF-κB and Ras-Raf-MEK-ERK-CREB signaling (red arrows), contributing to COX2 transcription and as a consequence prostaglandins production in the U.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay, Membrane, Transduction, Activation Assay, Protein-Protein interactions, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: The experimental procedure consists of two steps. Step 1 involved the investigation of IL1B and IL1R1 gene (RT-PCR) and protein (IHC) expression in reproductive tissues (T, P, and SV) in six male rabbits. Protein IL1B evaluation (WB) in the seminal plasma of 5 rabbits was included in this step. In Step 2, the U of six rabbits sliced into small pieces was incubated in vitro for two hours, and then the concentration of IL1B, NGF, PGE2, and PGF2α was assessed in the culture medium using ELISA method. The U (n = 6) evaluation of IL1R1 and IL1R3 gene (qPCR) and IL1R1 protein (IHC) were included in this step.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Clinical Proteomics, Incubation, In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: PCR primer. The table lists the gene name (acronym), the sequence identification number, the forward (F) and reverse (R) primers, and the amplicon length in the base pair (bp).

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques: Sequencing, Amplification

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: Antisera characteristics and dilutions. The table indicates the antibody name, the species in which the antibody was raised, and the working solution.

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation

doi: 10.3390/ijms252010986

Figure Lengend Snippet: Plate well design. The table shows the EGs from 1 to 7 (first column on the left) and the corresponding ligands (IL1B and NGF), or receptor antagonists (IL1R1) or inhibitors (TRKA, p75NTR, and COX inhibitors) used in the respective EG (when in wells added as +). Each column indicates the working solution and the reagent provider (as a note).

Article Snippet: Human NGF and IL1B proteins (catalog reference DY256 and E-EL-RB0013, respectively) were analyzed using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques:

Journal: medRxiv

Article Title: ACK1 and BRK non-receptor tyrosine kinase deficiencies are associated with familial systemic lupus and involved in efferocytosis

doi: 10.1101/2024.02.15.24302255

Figure Lengend Snippet: Cell lysates from 293T cells were incubated with anti-AKT. Immunoprecipitated proteins were probed with anti-phosphotyrosine and anti-AKT antibodies. (B) Western blot analysis for STAT3 phosphorylation by ACK1 and BRK . Lysates from 293T cells coexpressing STAT3 and Flag-tagged WT or mutant forms (A156T and K161Q) of ACK1 or mutant forms (G257A and G321R) of BRK were probed with anti-phospho-STAT3 (Tyr705), anti-STAT3 and anti-Flag antibodies. For analysis of BRK, cells were treated with 100 ng/ml EGF for 10 min. (C) RAC activation by WT ACK1 and BRK. Cell lysates from 293T cells expressing WT or mutant forms of ACK1 ( left ) and lysates from 293T cells expressing WT or mutant forms of BRK ( right ) were incubated with GST-PAK CRIB sepharose beads, and the level of RAC1 GTP was determined by immunoblotting with anti-Rac1 antibody. Lysates were also probed with anti-Rac1, anti-FLAG and anti-tubulin antibody. For analysis of BRK, 293T cells were cotransfected with CAS and stimulated with 100 ng/ml EGF for 10 min. ( D) MERTK increases kinase activity of BRK and ACK1. IP kinase assay. ACK1 (left) was immunoprecipitated from 293T cells co-transfected with Flag-tagged ACK1 WT, ACK1 A156T, or ACK1 K161Q and MERTK with anti-Flag Ab. Immunoprecipitated proteins were used in duplicate in vitro for kinase reactions with WASP synthetic peptide and results represented as pmol phosphate transferred. BRK (right) was immunoprecipitated as above from 293T cells co-transfected with Flag-tagged BRK WT or mutants and MERTK with anti-Flag Ab. Kinase reactions was performed with peptide AEEEIYGEFEAKKKG, and represented as above. P-values were calculated using an Anova test (Tukey’s multiple comparison test) (p > 0.05 (ns); p < 0.05 (*); p < 0.01 (**); p < 0.001 (***); p < 0.0001 (****). (E ) Regulation of inflammatory response. Significant normalized enrichment scores (NES) for GO ‘positive regulation of acute inflammation’ gene set, GO ‘negative regulation of inflammatory response’ gene set, and GO ‘AKT_UP.V1_UP’ gene set in WT and mutant macrophages, and WT treated with AIM100 (2 µM) or Cpd4f (0.5 µM), exposed to apoptotic cells, with 3 replicates per experimental condition. Significant enrichment (p-value<0.05 and FDR (q-value) <0.25) are calculated as reported in methods. (F) Table of the top 10 differentially regulated genes by apoptotic cells in WT macrophages are not differentially expressed in mutant macrophages and WT macrophages treated with AIM100 or Cpd4f (treated as in E). Numbers indicate FDR (q-value). Known target genes of STAT3 and AKT are labeled in blue and red respectively ( G ) TNF mRNA production by WT macrophages treated with AIM100 (2 µM) 4hrs after exposure to apoptotic cells. n=6, from 2 independent experiments. ( H,I) TNF and IL1b production by macrophages , as measured by ELISA on media collected from mutant and isogenic WT macrophages (C12.1) incubated with mouse apoptotic thymocytes for 90 min, then stimulated with LPS (1 ng/ml) for 18 h. n≥4, from ≥2 independent experiments. P-values in G-I were calculated using an Anova test with Tukey’s correction for multiple comparisons.

Article Snippet: ELISA was conducted according to manufacturer’s protocol (Human TNF-alpha DuoSet (R&D; DY210) and IL1b DuoSet (R&D, DY201) ELISA kit).

Techniques: Incubation, Immunoprecipitation, Western Blot, Mutagenesis, Activation Assay, Expressing, Activity Assay, IP-Kinase Assay, Transfection, In Vitro, Comparison, Labeling, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Oncology

Article Title: IL1 Pathway in HPV-Negative HNSCC Cells Is an Indicator of Radioresistance After Photon and Carbon Ion Irradiation Without Functional Involvement

doi: 10.3389/fonc.2022.878675

Figure Lengend Snippet: Evaluation of SASP gene expression after irradiation with 8 Gy photons or 4 Gy 12 C-ions. (A) Schematic of experimental procedure. (B) Principal component analysis (PCA) on z-scaled data of the five cell lines for the 49 SASP factors. (C) Hierarchical clustering (fold change >1.5) for 11 genes to separate irradiated from unirradiated samples for photons and (D) 12 C-ions. (E) Plot depicting fold change and q value for significantly differently expressed genes between unirradiated and irradiated samples for photons and 12 C-ions. (F) IL1A and (G) IL1B values 72 h after irradiation were calibrated to the unirradiated control and correlated with radioresistance AUC or senescence AUC. (H) Correlation of IL1 pathway activity with radioresistance AUC or senescence AUC. Experiments were performed in three biol. replicates. Values are MV+/- SEM. T-test statistics were used for p value calculation. p < 0.05 are considered significant. R 2 : Pearson coefficient. For PCA and hierarchical clustering, log2-transformed, normalized (mean = 0, var = 1) data was used. Benjamini–Hochberg correction was applied, and q < 0.05 was considered significant. (I) Detection of IL1B protein secretion by ELISA up to 6 days after irradiation with 8 Gy photons in the cell lines UPCI:SCC131, UPCI:SCC040, and Cal33.

Article Snippet: Cytokine concentrations in cell culture supernatants were measured with the human IL1B DuoSet ELISA System (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Gene Expression, Irradiation, Control, Activity Assay, Transformation Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Oncology

Article Title: IL1 Pathway in HPV-Negative HNSCC Cells Is an Indicator of Radioresistance After Photon and Carbon Ion Irradiation Without Functional Involvement

doi: 10.3389/fonc.2022.878675

Figure Lengend Snippet: Cell survival after siRNA knockdown of ILA or/and IL1B. (A) Colony formation assay was initiated 1 day after transfection. Knockdown and control cells were irradiated with 2–8 Gy photons or 1–3 Gy 12 C-ions. (B) Foci formation in response to 2-Gy photons or 1-Gy 12 C-ions was examined by γH2AX/53BP1 immunofluorescence 24 h after irradiation. (C) Flow cytometry-based analysis of senescence after siRNA knockdown of IL1B in the cell line UPCI:SCC040. IL1B knockdown and control samples were irradiated with 6 Gy photon or 3 Gy 12 C-ion, and senescence was determined from days 2 to 6 after irradiation. Experiments were performed in triplicates. MV+/- SEM are given. p values were calculated using t-test statistics. p = ns describes no significance was detected.

Article Snippet: Cytokine concentrations in cell culture supernatants were measured with the human IL1B DuoSet ELISA System (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Knockdown, Colony Assay, Transfection, Control, Irradiation, Immunofluorescence, Flow Cytometry