recombinant il 24  (MedChemExpress)

Journal: Computational and Structural Biotechnology Journal

Article Title: seneR: An R package for comprehensive senescence assessment and its application in type 2 diabetes and osteoarthritis

doi: 10.1016/j.csbj.2025.12.031

Figure Lengend Snippet: Verification of the therapeutic effect of PM in delaying the senescence of chondrocytes. (a) Schematic diagram of primary chondrocyte isolation and in vitro experimental design. (b) Representative images of brightfield, Alcian blue, Safranin O, and Toluidine blue staining of chondrocytes. (c, d) Protein levels of p16 and SLPI detected by Western blot (WB) after IL-1β treatment (c) or Slpi overexpression (OE-Slpi; d). (e, f) Representative images of SA-β-gal staining (e) and quantitative analysis of positive cells (f) in OE-negative control (OE-NC) or OE-Slpi chondrocytes. (g, h) Representative images of SA-β-gal staining (g) and quantitative analysis of positive cells (h) in chondrocytes treated with 10 ng/ml IL-1β alone or combined with 1 μM PM. (i, j) Quantitative real-time PCR (qPCR) analysis of p16 and MMP3 mRNA expression in chondrocytes treated with 10 ng/ml IL-1β alone or combined with 1 μM PM. (k, l) Representative images of SA-β-gal staining (k) and quantitative analysis of positive cells (l) in chondrocytes treated with H₂O₂ alone or combined with 1 μM PM. (m, n) qPCR analysis of p16 and MMP3 mRNA expression in chondrocytes treated with H₂O₂ alone or combined with 1 μM PM. Data are presented as mean ± standard deviation (n = 3 independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant. (Student’s t -test: f; ANOVA: h, i, j, l, m, n). OE, overexpression; NC, negative control.

Article Snippet: For over-expression SLPI, chondrocytes were transfected with pcDNA3.1/Slpi full-length plasmid using Lipo8000 (Beyotime) according to the manufacturer’s instructions.For IL-1β treatment, mouse primary chondrocytes were treated with recombinant mouse IL-1β (10 ng/ml, MedChemExpress) for 24 h. For H2O2-induced senescence, mouse primary chondrocytes were treated with H2O2 (200 μM, MKBio Shanghai) for 2 h. For PM treatment, mouse primary chondrocytes were treated with or without PM (MedChemExpress), dissolved in DMSO (Dimethyl sulfoxide, MedChemExpress).

Techniques: Isolation, In Vitro, Staining, Western Blot, Over Expression, Negative Control, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: International immunopharmacology

Article Title: House dust mite induced mucosal barrier dysfunction and type 2 inflammatory responses via the MAPK/AP-1/IL-24 Signaling pathway in allergic rhinitis.

doi: 10.1016/j.intimp.2024.113972

Figure Lengend Snippet: Fig. 4. Regulation of IL-24 by transcription factor AP-1. (A) RT-qPCR analysis of IL-24 mRNA expression levels in human nasal mucosal tissue samples (left, n=15) and ELISA detection of IL-24 levels in human serum samples (right, n=5). (B) Measurement of IL-24 mRNA in nasal mucosal and serum samples from HDM-sensitized AR mouse models (n=4). (C) Positive correlation between IL-24 expression and FOS and JUN levels in human nasal mucosal tissue samples. (D) Positive correlation between IL-24 expression and FOS and JUN levels in mouse nasal mucosal tissue samples. (E) Predicted AP-1 binding sites in the IL-24 promoter region and design of fragments for subsequent ChIP analysis. (F) AP-1 binding motif. (G) Confirmation of RT-qPCR products in ChIP assays using nucleic acid electrophoresis. (H) ChIP-seq data from the ENCODE database verifying AP-1 binding peaks on the promoter region of IL-24 in A549 cells. Error bars represent mean ± SD. Ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant human IL-24 was obtained from MedChemExpress.

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay, Nucleic Acid Electrophoresis, ChIP-sequencing

Journal: International immunopharmacology

Article Title: House dust mite induced mucosal barrier dysfunction and type 2 inflammatory responses via the MAPK/AP-1/IL-24 Signaling pathway in allergic rhinitis.

doi: 10.1016/j.intimp.2024.113972

Figure Lengend Snippet: Fig. 5. IL-24 downregulates occludin and ZO-1 via the JAK/STAT1/3 pathway. (A) Phosphorylated STAT1 and STAT3 expression in human nasal mucosal samples was assessed using Western blotting (n = 3). (B) Phosphorylated STAT1 and STAT3 expression in mouse nasal mucosal samples was also evaluated using Western blotting (n = 3). (C) IL-24 mRNA expression and increased IL-24 levels in the supernatant were measured in HNEpC cells stimulated with HDM solution for 24 h (n = 3). (D) HNEpC were treated with HDM solution for 24 h, followed by re-treatment with the supernatant for another 24 h. Western blot was used to assess phos phorylated STAT1/STAT3 and TJ proteins (n = 3). (E) The expression of TJ proteins occludin and ZO-1 was significantly reduced in the HDM-treated samples, indicating compromised epithelial integrity. (F) Cell permeability was evaluated for the samples in (D); the left panel shows TEER measurements, and the right panel presents FD4 permeability assays (n = 3). (G) HNEpC cells were treated with recombinant human IL-24 protein for 24 h, followed by Western blotting to measure phosphorylated STAT1 and STAT3, as well as TJ protein expression (n = 3). (H) Results revealed a decrease in the expression of TJ proteins occludin and ZO-1 in the samples treated as indicated in (G) (n = 3). (I) Cell permeability was assessed for the samples in (G); the left panel shows TEER measurements, and the right panel presents FD4 permeability assays (n = 3). (J) STAT1 was knocked down using si-STAT1, and HNEpC cells were treated with recombinant human IL-24 for 24 h. Phosphorylated STAT1 and tight junction (TJ) protein expression were then assessed by Western blotting (n = 3). (K) HNEpC cells were treated with the STAT1 inhibitor Fludarabine and recombinant human IL-24 for 24 h. Phosphorylated STAT1 and tight junction (TJ) protein expression were then measured by Western blotting (n = 3). Error bars represent mean ± SD. ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant human IL-24 was obtained from MedChemExpress.

Techniques: Expressing, Western Blot, Permeability, Recombinant

Journal: International immunopharmacology

Article Title: House dust mite induced mucosal barrier dysfunction and type 2 inflammatory responses via the MAPK/AP-1/IL-24 Signaling pathway in allergic rhinitis.

doi: 10.1016/j.intimp.2024.113972

Figure Lengend Snippet: Fig. 6. Blocking IL-24 protects the epithelial barrier. (A) After knockdown of IL20Rβ, HNEpC were treated with HDM solution for 24 h, resulting in a decrease in the mRNA expression of occludin and ZO-1. n = 3. (B) Western blot analysis of samples from (A) revealed the phosphorylation levels of STAT1/3 and the expression of TJ proteins. n = 3. (C) The cellular permeability of samples from (A) was assessed; the left panel shows TEER measurements, and the right panel displays FD4 permeability experiments. n = 3. (D) Schematic diagram of the AR disease model induction and stimulation in HDM-sensitized mice treated with Ruxolitinib. (E) The left panel shows the ELISA detection of serum IgE levels in mice, while the right panel presents the overall score of nasal symptoms. n = 8. (F) Representative images of H&E staining of mouse nasal mucosa, with red lines indicating the epithelial boundary of the nasal mucosa. (G) Measurement of nasal mucosal thickness in H&E- stained sections. n = 3. (H) Eosinophil counts in high-power fields of H&E-stained nasal mucosa. n = 3. (I) Western blot analysis of mouse nasal mucosa samples to assess the phosphorylation of STAT1/3 and the expression of TJ proteins. n = 3. (J) FD4 levels in NALF (n = 4) and serum (n = 8). (K) Representative images of immunofluorescence analysis of occludin and ZO-1 in mouse nasal mucosa tissue. n = 3. Error bars represent mean ± SD. ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant human IL-24 was obtained from MedChemExpress.

Techniques: Blocking Assay, Knockdown, Expressing, Western Blot, Phospho-proteomics, Permeability, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence

Journal: International immunopharmacology

Article Title: House dust mite induced mucosal barrier dysfunction and type 2 inflammatory responses via the MAPK/AP-1/IL-24 Signaling pathway in allergic rhinitis.

doi: 10.1016/j.intimp.2024.113972

Figure Lengend Snippet: Fig. 7. IL-24 promotes IL-33 production. (A) Treatment of HNEpC with HDM solution for 24 hours followed by a second 24-hour stimulation resulted in increased IL- 33 mRNA expression and IL-33 levels in the supernatant (n=3). (B) Treatment with recombinant human IL-24 for 24 hours also increased IL-33 mRNA expression and supernatant IL-33 levels in HNEpC (n=3). (C) After IL20Rβ knockdown, HDM treatment of HNEpC for 24 hours led to decreased IL-33 mRNA expression and IL-33 levels in the supernatant (n=3). (D) Following 2 hours of Ruxolitinib treatment, subsequent HDM stimulation of HNEpC for 24 hours resulted in reduced IL-33 mRNA expression and supernatant IL-33 levels (n=3). (E) Serum IL-33 levels and IL-33 mRNA expression in nasal mucosa samples from mice were measured (n=3). (F) A schematic representation showing how HDM upregulates IL-24, damaging the epithelial barrier. Error bars represent mean ± SD. ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant human IL-24 was obtained from MedChemExpress.

Techniques: Expressing, Recombinant, Knockdown