cd130  (Bioss)

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Exposure to IL-6 (10 ng/mL, 12 hours) induced increases in ODAM mRNA levels in Ca9-22 cells (A), HSY cells (B), and Sa3 cells (C), which were inhibited by SC144 (1 μM). ODAM and GAPDH mRNA levels were analyzed using real-time PCR. The experiments were performed in triplicate. Quantitative analyses of the data sets are presented with standard deviations. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin-6, GAPDH: glyceraldehyde 3-phosphate dehydrogenase, PCR: polymerase chain reaction. Significant differences from the control are indicated by ** P <0.01.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Control

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Protein levels were assessed using Western blotting following stimulation with IL-6 (10 ng/mL) for durations of 10 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, and 24 hours in Ca9-22 cells. Densitometric analyses of the Western blot results for ODAM, p-STAT3, STAT3, p-p65, p65, p-IKKα/β, IKKβ, and IKKα are presented. The intensity of the α-tubulin band was normalized to 1, and graphs were generated to illustrate the relative intensities of the other protein bands. IL-6: interleukin-6, ODAM: odontogenic ameloblast-associated protein, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, p65: nuclear factor-kappa B p65, p-p65: phosphorylated p65, IKKα: IκB kinase α, IKKβ: IκB kinase β, p-IKKα/β: phosphorylated IκB kinase α/β. The data represent the mean ± standard deviation from 3 independent experiments, with * P <0.05 and ** P <0.01 indicating statistical significance.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Western Blot, Generated, Standard Deviation

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: (A) LUC activity of −85ODAM, −116ODAM, −174ODAM, −200ODAM, −300ODAM, −330ODAM, −480ODAM, −700ODAM, and −950ODAM constructs were increased by IL-6 (10 ng/mL) after 12 hours of treatment in Ca9-22 cells. Transcriptional activity was determined from 4 separate transfections, and the combined values are expressed with SDs. (B) Effects of kinase inhibitors on the LUC activity of −480ODAM, stimulated by IL-6. Transient transfection analyses of −480ODAM in Ca9-22 cells, with or without IL-6 (10 ng/mL) treatment, were conducted alongside the effects of inhibitors of PKC (H7, 5 μM), PKA (KT5720, 100 nM), tyrosine kinase (HA, 1 μM), MEK1/2 (U0126, 5 μM), PI3K (LY294002, 10 μM), NF-κB (triptolide, 100 nM), STAT3 (stattic, 10 µM), STAT1 (fludarabine, 10 µM), and gp130 (SC144, 1 μM). Transcriptional activity was determined from 3 separate transfections, with the combined values expressed with SDs. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin-6, LUC: luciferase, SD: standard deviation, PKC: protein kinase C, PKA: protein kinase A, MEK1/2: mitogen-activated protein kinase kinase 1/2, PI3K: phosphatidylinositol 3-kinase, NF-κB: nuclear factor-kappa B, STAT: signal transducer and activator of transcription, gp130: glycoprotein 130. Significant differences from the control are indicated by ** P <0.01.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Activity Assay, Construct, Transfection, Luciferase, Standard Deviation, Control

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Supershift gel shift assays were performed using anti-YY1, C/EBPβ, GATA, STAT3, and p-STAT3 antibodies for each reaction. The DNA-protein complexes were then separated by electrophoresis on a 6% polyacrylamide gel and visualized using the ChemiDoc Touch MP Imaging System. YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Gel Shift, Electrophoresis, Imaging, Binding Assay

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: (A) PCR bands corresponding to DNA-protein complexes immunoprecipitated with antibodies indicated that YY1, C/EBPβ, GATA, STAT3, and p-STAT3 interact with chromatin fragments containing the YY1, C/EBP, GATA, and GATE1-3 sequences. These interactions were increased in Ca9-22 cells following stimulation with IL-6 (10 ng/mL). Input DNA served as a control in the PCR analysis. (B) ChIP analyses were conducted to assess the binding of transcription factors to YY1, C/EBP, GATA, and GATE1-3 in the presence of IL-6 and various kinase inhibitors. Treatment with HA, U0126, LY294002, and triptolide almost completely abolished, and KT5720 partially inhibited, the IL-6-induced (10 ng/mL, 12 hours) binding of YY1, C/EBPβ, and GATA transcription factors to their respective DNA sequences. Additionally, treatment with stattic almost completely abolished, while fludarabine did not inhibit, the IL-6-induced binding of STAT3 transcription factors to GATE1-3. ChIP: chromatin immunoprecipitation, YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, GATE1-3: interferon-γ activated transcriptional element sites 1-3, ODAM: odontogenic ameloblast-associated protein, PCR: polymerase chain reaction, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, IL-6: interleukin-6.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Immunoprecipitation, Control, Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: IL-6–regulated ODAM gene expression is supported by protein–DNA interactions at the YY1, C/EBP, GATA, and GATE1-3. The signaling pathway involves gp130, JAK, STAT3, p-STAT3, PI3K, AKT, NF-κB, GPCR, SOCS3, SHP2, ERK, Grb2, Shc, Ras, and Raf. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin 6, YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, GATE1-3: interferon-γ activated transcriptional element 1-3, gp130: glycoprotein 130, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, PI3K: phosphatidylinositol 3-kinase, NF-κB: nuclear factor-kappa B, GPCR: G protein-coupled receptor.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Gene Expression, Binding Assay

Journal: Cell reports

Article Title: A basophil-fibroblast pro-inflammatory axis fuels type 2 skin inflammation

doi: 10.1016/j.celrep.2025.116114

Figure Lengend Snippet: (A) Experimental outline of Il4ra fl/fl vs. Il4ra ΔPdgfra mice treated with vehicle or SC144 during MC903. (B) Representative IF staining of pSTAT3 (red) and PDGFRA (green) in Il4ra fl/fl and Il4ra ΔPdgfra with vehicle or SC144 in d7 MC903, quantified at right. * p < 0.05, ** p < 0.01, one-way ANOVA with Dunn’s multiple comparisons test. (C) Representative gross clinical images and H&E from Il4ra fl/fl vs. Il4ra ΔPdgfra mice with vehicle or SC144 at d7 MC903. (D) Skin thickness measurements of Il4ra fl/fl vs. Il4ra ΔPdgfra mice with vehicle or SC144 at d7 MC903. Data are mean ± S.E.M. * p < 0.05, *** p < 0.001, one-way ANOVA with Tukey’s multiple comparisons test. (E) Skin thickness quantification of Il4ra fl/fl vs. Il4ra ΔPdgfra mice with vehicle or SC144 at d7 MC903. Data are mean ± S.E.M. * p < 0.05, *** p < 0.001, one-way ANOVA with Tukey’s multiple comparisons test. (F) CD45 + proportion in Il4ra fl/fl vs. Il4ra ΔPdgfra mice at d7 MC903. Data are mean ± S.E.M. * p < 0.05, ** p < 0.01, one-way ANOVA with Tukey’s multiple comparisons test. (G) Proposed model of basophil-induced OSM and IL-4/13 synergistic stimulation of pro-inflammatory fibroblasts that recruit immune cells to promote inflammation.

Article Snippet: SC144 gp130 inhibitor , MedChemExpress , HY-15614.

Techniques: Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: IL-6 signaling is required for the development and regeneration of ear cartilage in microtia

doi: 10.3389/fcell.2025.1625058

Figure Lengend Snippet: IL-6 binds to IL-6R and GP130 to activate the JAK/STAT3 and PI3K/AKT pathways in primary chondrocytes and PSCECs. (A, B) IP-MS identified receptors that bind to IL-6 among the membrane proteins of primary chondrocytes and PSCECs, and potential binding possibilities between IL-6, IL-6R, and GP130 were discovered. (C) IF assay showing the colocalization of IL-6, IL-6R, and GP130 in primary chondrocytes and PSCECs. Scale bar, 50 μm. (D) Co-IP confirmed the protein-binding relationship among IL-6, IL-6R, and GP130 in primary chondrocytes and PSCECs. (E) RNA-seq of primary chondrocytes and PSCECs detected the expression of cartilage fibrosis-related genes after IL-6 (50 ng/mL) stimulation. (F) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the upregulated genes in primary chondrocytes and PSCECs after IL-6 stimulation showcased the signaling pathways involved in these genes. (G) GO enrichment analysis of the upregulated genes in primary chondrocytes and PSCECs after IL-6 stimulation. (H) GSEA revealed that the gene terms for extracellular collagen, ear morphogenesis, pattern cognitive receptor activation, and ear development were highly enriched in the IL-6 treatment group of primary chondrocytes and PSCECs. (I) Western blots showing the protein levels of SOX9, COL1A1, Collagen II and JAK/STAT3 and PI3K/AKT signaling pathways in different groups; GAPDH was used as a control.

Article Snippet: Primary antibodies against IL-6 (1:300, Proteintech, Chicago, United States), IL-6R (Abcam, 1:1,000, Cambridge, UK), GP130 (Proteintech, 1:1,000, Chicago, United States), CD90 (Abcam, 1:600, Cambridgeshire, UK), and SOX9 (Abcam, 1:1,000, Cambridge, UK) were used to incubate the slices at 4°C overnight.

Techniques: Protein-Protein interactions, Membrane, Binding Assay, Co-Immunoprecipitation Assay, Protein Binding, RNA Sequencing, Expressing, Activation Assay, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: IL-6 signaling is required for the development and regeneration of ear cartilage in microtia

doi: 10.3389/fcell.2025.1625058

Figure Lengend Snippet: IL-6 signaling participates in the intercellular communication network between PSCECs and primary chondrocytes. (A) The cell co-culture system using the transwell plate with a pore size of 0.4 μm. (B) ELISA showed the change of IL-6 level in the cell culture medium after co-cultured with PSCECs. (C) The proliferative ability of PSCECs and primary chondrocytes was determined by CCK8 assay in different groups. (D) The DNA synthesis of PSCECs and primary chondrocytes was detected by EdU assay. Scale bar, 50 μm. (E) The migrative capacity of primary chondrocytes and PSCECs in different groups was assessed by transwell assay. Scale bar, 50 μm. (F) The migration of primary chondrocytes and PSCECs in different groups was detected by scratch wound assay. Scale bar, 100 μm. (G) Western blots identified the protein level of SOX9, COL1A1, Collagen II and JAK/STAT3 and PI3K/AKT components pathway in different groups, GAPDH was used as a control. (H) In microtia, mIHC assay showed that IL-6, IL-6R and GP130 co-expressed in CD90 + stem cells and SOX9+ chondrocytes. Scale bar, 50 μm *P < 0.05, **P < 0.01.

Article Snippet: Primary antibodies against IL-6 (1:300, Proteintech, Chicago, United States), IL-6R (Abcam, 1:1,000, Cambridge, UK), GP130 (Proteintech, 1:1,000, Chicago, United States), CD90 (Abcam, 1:600, Cambridgeshire, UK), and SOX9 (Abcam, 1:1,000, Cambridge, UK) were used to incubate the slices at 4°C overnight.

Techniques: Co-Culture Assay, Pore Size, Enzyme-linked Immunosorbent Assay, Cell Culture, CCK-8 Assay, DNA Synthesis, EdU Assay, Transwell Assay, Migration, Scratch Wound Assay Assay, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: IL-6 signaling is required for the development and regeneration of ear cartilage in microtia

doi: 10.3389/fcell.2025.1625058

Figure Lengend Snippet: IL-6 promotes the development of embryonic ear in vivo . (A) We used tretinoin to construct the animal model of microtia, and we used AAV-IL-6 for gene therapy. (B) Representative images of pregnant mice and their fetuses after birth. (C) The area of the fetal mice ear in different group. (D) The liver and kidney function in pregnant mice. (E) The HE staining of fetal mice ear. Scale bar, 50 μm. (F) The Alcian Blue and Saffron-O staining of fetal mice ear. Scale bar, 50 μm. (G) The frozen section was used to detect GFP in fetal mice ear. Scale bar, 50 μm. (H) The protein level of IL-6 was detected in fetal mice ear by using IHC. Scale bar, 50 and 20 μm. (I) The protein level of IL-6 was detected in fetal mice ear by using Western blots, β-was used as a control. (J) mIHC assay showed that IL-6, IL-6R and GP130 co-expressed in CD90 + cells and SOX9+ cells in the ears of fetal mice. Scale bar, 50 μm *P < 0.05, **P < 0.01.

Article Snippet: Primary antibodies against IL-6 (1:300, Proteintech, Chicago, United States), IL-6R (Abcam, 1:1,000, Cambridge, UK), GP130 (Proteintech, 1:1,000, Chicago, United States), CD90 (Abcam, 1:600, Cambridgeshire, UK), and SOX9 (Abcam, 1:1,000, Cambridge, UK) were used to incubate the slices at 4°C overnight.

Techniques: In Vivo, Construct, Animal Model, Staining, Western Blot, Control