Journal: Journal of Molecular Histology

Article Title: Interleukin-1 Receptor-Associated Kinase 3 Attenuates Chondrocyte Senescence and Osteoarthritis via Inhibition of the TLR7/9–NF-κB Axis

doi: 10.1007/s10735-026-10804-4

Figure Lengend Snippet: illustrates the role of IRAK3 in modulating TLR7/9 signaling and reducing inflammation in chondrocytes. AT791 is a TLR7 and TLR9 inhibitor. The treatment concentration is 3 µM and the treatment time is 48 h. The TLR9 agonist used is ODN 1826, with a concentration of 1 µg/ml and a treatment time of 48 h. The TLR7 agonist used is Resiquimod, with a concentration of 1 µM and a treatment time of 48 h. Both were purchased from MedChemExpress, HY-124,603, HY-146,245 and HY-13,740. Panels A and B show Western blot analysis of TLR7 and TLR9 expression following IRAK3 knockdown and overexpression, respectively. Panels C through E present ELISA results indicating levels of IL-1β, IL-6, and TNF-α after IRAK3 knockdown. Similarly, panels F through H display ELISA findings for IL-1β, IL-6, and TNF-α levels following IRAK3 overexpression. In panel I, chondrocytes were treated with TLR7/9 inhibitors and agonists, and β-galactosidase staining was performed on senescent chondrocytes for statistical analysis. (* p < 0.05, ** p < 0.01)

Article Snippet: The treatment concentration is 3 μM and the treatment time is 48 h. The TLR9 agonist used is ODN 1826, with a concentration of 1 μg/ml and a treatment time of 48 h. The TLR7 agonist used is Resiquimod, with a concentration of 1 μM and a treatment time of 48 h. Both were purchased from MedChemExpress, HY-124,603, HY-146,245 and HY-13,740.

Techniques: Concentration Assay, Western Blot, Expressing, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Staining

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Figure Lengend Snippet: Figure 4 | Dendritic cell uptake of MUS/OT NPs correlates with effective vaccine response. (a) C57Bl/6 mice (n ¼ 5 mice per group) were injected s.c. with 300 mg MUS/OT or PEG AuNPs, followed by ICP-AES quantification of total gold NP accumulation in lumbar, inguinal, axillary LNs 24 h later. Shown are mean±s.d. ***Po0.001 by unpaired t-tests. (b,c) C57Bl/6 mice (n ¼ 3 per group) were injected s.c. with 100 mg MUS/OT, MPSA or PEG AuNPs, and lymph nodes were excised, digested and stained for CyTOF2 analysis 24 h later. Shown are representative Au histograms showing AuNP levels in various leukocyte populations (b) and mean number of NPs per cell for lymph node cell populations (c). (d) Schematic structure of SIINFEKL peptide construct and coupling to MUS/OT NP surface. (e–i) C57Bl/6 mice (n ¼ 5 per group) were immunized s.c. on days 1 and 14 with 8 mg CpG mixed with SIINFEKL- conjugated MUS/OT NPs (10 mg peptide), 50 mg SIINFEKL peptide, 10 mg SIINFEKL peptide or 10 mg SIINFEKL peptide construct. Animals were then challenged with 2.5 105 B16-OVA tumour cells s.c. in the flank on day 150. Shown are representative flow cytometry plots of SIINFEKL tetramer staining (e) and mean SIINFEKL tetramer þ CD8 þ T cells in blood on day 29 (f), representative intracellular cytokine staining flow cytometry (g) and mean percentages of cytokine þ T cells (h) 6 days after tumour challenge, and tumour size measurements over time (i). Shown are means±s.e.m. *Po0.05; **Po0.01, ***Po0.001 by one-way ANOVA with Bonferroni post tests.

Article Snippet: Eight-week-old female C57BL/6 mice were immunized (primed on day 1, boosted on day 14) with 8 mg of CpG (ODN 1826 VacciGrade, InvivoGen) mixed with SIINFEKL peptide (10mg peptide-conjugated AuNP, 10 mg free peptide, 50mg free peptide or 10mg free peptide construct).

Techniques: Injection, Staining, Construct, Cytometry