Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Significant Ups genes in the acetabular cartilage of the rats with DDH, as determined by mRNA-seq analysis. (A) Neonatal rats were subjected to straight-leg swaddling for 10 days. A total of 4 weeks later, (a) morphology, (b) H&E staining and (c) SO/FG staining were assessed in the hip joints of the rats. (B) Immunohistochemical staining and (E) semi-quantitative analysis of COL2A1 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (C) Relative gene expression levels of COL2A1, IL-1β and IL-6 in the acetabular cartilage of the 4-week-old control rats and rats with DDH, as determined by reverse transcription-quantitative PCR. (D) Levels of IL-6 in the supernatants of acetabular cartilage from the 4-week-old rats with DDH, as determined by ELISA. (F) Significant Ups in the acetabular cartilage of rats with DDH were identified by mRNA-seq analysis with the screening criteria of log 2 FC >4 and P.adj<0.01. Principal component analysis results and a heatmap of these significant Ups are shown. (G) GO enrichment analysis of significant Ups in the acetabular cartilage of the rats with DDH. Data are presented as the mean ± SD. *** P<0.001. (C-E) Representative results of four independent experiments are shown. Ups, upregulated genes; BP, biological process; CC, cellular component; COL2A1, type II collagen; DDH, developmental dysplasia of the hip; FC, fold change; GO, Gene Ontology; H&E, hematoxylin and eosin; IHC, immunohistochemistry; IL, interleukin; IOD, integrated optical density; MF, molecular function; mRNA-seq, mRNA sequencing; SO/FG, Safranin O/Fast Green.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Staining, Immunohistochemical staining, Control, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Sequencing

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 expression is upregulated in IL-1β-induced chondrocytes. (A) Schematic diagram showing the experimental protocol in vitro . Primary chondrocytes isolated from 5-day-old rats were subjected to treatment with 10 ng/ml IL-1β. The protein and mRNA expression levels of DUSP26 in chondrocytes were detected by RT-qPCR and western blotting. (B) Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the protein and mRNA expression levels of DUSP26 in the chondrocytes were detected by RT-qPCR and western blotting. (C) Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the protein and mRNA expression levels of DUSP26 in the chondrocytes were assessed by RT-qPCR and western blotting. Data are presented as the mean ± SD. ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. Ad-EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Expressing, In Vitro, Isolation, Quantitative RT-PCR, Western Blot, Over Expression, Construct, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 overexpression alleviates IL-1β-induced chondrocyte injury. Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (A) Expression levels of COL2A1 and COL1A1 in the chondrocytes were detected by RT-qPCR and western blotting. (B) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (C) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were determined by ELISA. Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (D) Expression levels of COL2A1 and COL1A1 in the chondrocytes were evaluated by RT-qPCR and western blotting. (E) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (F) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were detected by ELISA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. COL1A1, type I collagen; COL2A1, type II collagen; EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Over Expression, Construct, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: HDAC1, HDAC2 and HDAC8 are DUSP26-interacting proteins in chondrocytes. (A) IP/MS analysis in DUSP26-overexpressing chondrocytes under IL-1β stimulation. (B) Flow chart of target protein screening. (C) Immunohistochemical staining of HDAC1, HDAC2 and HDAC8 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (D) Protein expression levels of HDAC1, p-HDAC1, HDAC2, p-HDAC2, HDAC8 and p-HDAC8 in the chondrocytes by western blotting. (E) Co-IP experiments with DUSP26 protein from extracts of IL-1β-treated chondrocytes followed by western blotting with indicated antibodies. DEG, differentially expressed gene; DUSP26, dual-specificity phosphatase 26; HDAC, histone deacetylase; IB, immunoblot; IL, interleukin; IP, immunoprecipitation; LC-MS/MS, liquid chromatography-tandem MS; mRNA-seq, mRNA sequencing; MS, mass spectrometry; p-, phosphorylated.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Protein-Protein interactions, Immunohistochemical staining, Staining, Control, Expressing, Western Blot, Co-Immunoprecipitation Assay, Histone Deacetylase Assay, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Sequencing, Mass Spectrometry

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Inactivation of HDAC1/2/8 inhibits DUSP26 silencing-triggered cartilage degeneration. (A) Total protein and phosphorylation levels of HDAC1, HDAC2 and HDAC8 in IL-1β-treated chondrocytes determined by western blotting. (B) Relative mRNA expression levels of HDAC1, HDAC2 and HDAC8 in chondrocytes, as determined by reverse transcription-quantitative PCR. (C) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (D) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. (E) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (F) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. (B-D) Representative results of three independent experiments are shown. COL1A1, type I collagen; DUSP26, dual-specificity phosphatase 26; HDAC, histone deacetylase; IL, interleukin; NC, negative control; ns, no significance; sh, short hairpin; TSA, trichostatin A.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Phospho-proteomics, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Negative Control