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: These antibodies included those specific for DUSP26 (cat. no. bs-7910R; BIOSS), COL2A1 (cat. no. AF0135; Affinity Biosciences), histone deacetylase (HDAC)1 (cat. no. AF6433; Affinity Biosciences), phosphorylated (p)-HDAC1 (Ser421) (cat. no. PA5-36810; Thermo Fisher Scientific, Inc.), HDAC2 (cat. no. AF6470; Affinity Biosciences), p-HDAC2 (Ser394) (cat. no. bs-5389R; BIOSS), HDAC8 (cat. no. AF6481; Affinity Biosciences) and p-HDAC8 (Ser39) (cat. no. AF3481; Affinity Biosciences).

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: These antibodies included those specific for DUSP26 (cat. no. bs-7910R; BIOSS), COL2A1 (cat. no. AF0135; Affinity Biosciences), histone deacetylase (HDAC)1 (cat. no. AF6433; Affinity Biosciences), phosphorylated (p)-HDAC1 (Ser421) (cat. no. PA5-36810; Thermo Fisher Scientific, Inc.), HDAC2 (cat. no. AF6470; Affinity Biosciences), p-HDAC2 (Ser394) (cat. no. bs-5389R; BIOSS), HDAC8 (cat. no. AF6481; Affinity Biosciences) and p-HDAC8 (Ser39) (cat. no. AF3481; Affinity Biosciences).

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

Journal: bioRxiv

Article Title: Cancer-associated KBTBD4 mutations induce differentiation defects and confer a unique therapeutic vulnerability

doi: 10.64898/2026.03.12.711277

Figure Lengend Snippet: (A) Volcano plot showing global protein expression in HL60 cells transduced with P311PP mutant. Blue and red dots show significantly decreased or increased proteins (adjusted p-value<0.05). CoREST components RREB1, RCOR1, KDM1A, RCOR3, and ZNF217 are highlighted in the volcano plot. The global proteome data was plotted after excluding PCDHGA11. (B) High throughput compound screening in HL60 cells stably expressing RCOR1-GFP treated with UM171 (200nM). Schematic of the screening rationale (top panel). The compounds were added at a final concentration of 1µM, and flow analysis was performed after 3 hours post treatment to measure the relative GFP expression (bottom panel). Red dots show the hits from the screen and the table on right-side shows the functional classification of the hits. (C) A dose-titration experiment showing the rescue of RCOR1-GFP by two class I specific HDAC inhibitors (mocetinostat and romidepsin) and three Pan HDAC inhibitors (belinostat, pracinostat and vorinostat). GFP mean fluorescence intensity of DMSO treatment was used as controls. Data from 3 replicates from 1 of 2 independent experiments with similar results are shown. (D) RCOR1 protein levels in P311PP expressing HL60 cells either treated with DMSO, belinostat (320nM) or mocetinostat (300nM) for 3 hours. (E) RCOR1 ELM2-GFP clones were expressed in HL60 cells and analyzed for the interaction with HDAC2 through immunoprecipitation. The degradation profiles of the corresponding alanine substitution clones to UM171 treatment are represented as a heat map. (F) Schematic representation of the mechanistic basis of HDAC inhibitors in preventing the CoREST degradation.

Article Snippet: The proteins were transferred to iBlot 2 PVDF membranes (#IB24001, Thermo Fisher Scientific) and probed with the following primary antibodies: RCOR1 (#14567), KDM1A (LSD1, #2184S), HDAC2 (#5113T, all from Cell Signaling Technology), GFP (#ab290, Abcam), and Actin (#612656, BD Biosciences).

Techniques: Expressing, Transduction, Mutagenesis, High Throughput Screening Assay, Stable Transfection, Concentration Assay, Functional Assay, Titration, Fluorescence, Clone Assay, Immunoprecipitation