Journal: The Journal of Biological Chemistry

Article Title: Piezo1 accelerates osteoarthritis progression via promotion of HBB-dependent oxidative phosphorylation

doi: 10.1016/j.jbc.2026.113489

Figure Lengend Snippet: Piezo1/IL-1β Synergy Drives OA and Metabolic Reprogramming. A , Representative Western blot bands of Piezo1, COL II, P16 ink4a and MMP13 in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h. B , quantitative analysis of Piezo1, COL II, P16 ink4a and MMP13 in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h ns, not significant, ∗ p < 0.05 and ∗∗ p < 0.01, n = 3. C , representative β-Galactosidase staining of the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h. D , quantitative analysis of β-Galactosidase-positive cell percentage of the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h ns, not significant, ∗∗ p < 0.01 and ∗∗∗ p < 0.001, n = 3. Scale bar: 100 μm. E , representative Western blot bands of HIF-1α, ATP5A1, LDHA and SOD2 in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h. F , quantitative analysis of HIF-1α, ATP5A1, LDHA and SOD2 in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with or without IL-1β and/or Yoda1 for 24 h ns, not significant, ∗ p < 0.05 and ∗∗ p < 0.01, n = 3. G , representative oxygen consumption rate (OCR) profiles and quantitative analysis of mitochondrial respiration parameters, including basal respiration, ATP production, maximal respiration, and spare respiratory capacity in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with IL-1β and Yoda1 for 24 h ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001, n = 3. H , Representative extracellular acidification rate (ECAR) profiles and quantitative analysis of glycolytic parameters, including glycolysis, glycolytic capacity, glycolytic reserve, and non-glycolytic acidification in the primary chondrocytes transfected with either empty vector (PcDNA3.1-IRES-eGFP) or mPiezo1-overexpression vector after treatment with IL-1β and Yoda1 for 24 h ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001, n = 3.

Article Snippet: The membranes were blotted with primary antibodies recognizing LDHA, p16 ink4a , ATP5A1, HIF-1α, HSP90 (diluted 1:20,000, Proteintech, Catalog: 60318-1-Ig), β-actin (diluted 1:50,000, Proteintech, Catalog: 66009-1-Ig), HBB, MMP13 (diluted 1:2000, Proteintech, Catalog: 18165-1-AP), SOD2 (diluted 1:20,000, Proteintech, Catalog: 24127-1-AP) and Collagen Type II.

Techniques: Western Blot, Transfection, Plasmid Preparation, Over Expression, Analysis, Staining

Journal: The Journal of Biological Chemistry

Article Title: Piezo1 accelerates osteoarthritis progression via promotion of HBB-dependent oxidative phosphorylation

doi: 10.1016/j.jbc.2026.113489

Figure Lengend Snippet: HBB Knockdown Rescues Mitochondrial Dysfunction. A , Representative Western blot bands and quantitative analysis of COL II, MMP13, P16 ink4a , ATP5A1, LDHA, HIF-1α and SOD2 in the primary chondrocytes transfected by mPiezo1-overexpression vector after treatment with siNC or siHBB in the presence of IL-1β and Yoda1 at 6 and 24 h. B , quantitative analysis of COL II, MMP13, P16 ink4a , ATP5A1, LDHA, HIF-1α, and SOD2 in the primary chondrocytes transfected by mPiezo1-overexpression vector after treatment with siNC or siHBB in the presence of IL-1β and Yoda1 at 6 {i} and 24 {ii} h. ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001, n = 3. C , representative β-Galactosidase Staining of the primary chondrocytes transfected by mPiezo1-overexpression vector after treatment with siNC or siHBB in the presence of IL-1β and Yoda1 at 24 h ∗∗∗ p < 0.001, n = 3. Scale bar: 20 μm. D , representative fluorescence images showing mitochondrial superoxide levels of the primary chondrocytes transfected by mPiezo1-overexpression vector after treatment with siNC or siHBB in the presence of IL-1β and Yoda1 at the indicated time points. Scale bar: 100 μm. E , quantification of the relative fluorescence unit in ( D ), ∗ p < 0.05 and ∗∗∗∗ p < 0.0001, n = 3. Scale bar: 100 μm. F , representative fluorescence images of TMRM staining in the primary chondrocytes transfected by mPiezo1-overexpression vector after treatment with siNC or siHBB in the presence of IL-1β and Yoda1 at the indicated time points. Scale bar: 100 μm. G , quantitative analysis of the relative fluorescence unit in ( E ), ∗ p < 0.05 and ∗∗∗∗ p < 0.0001, n = 3.

Article Snippet: The membranes were blotted with primary antibodies recognizing LDHA, p16 ink4a , ATP5A1, HIF-1α, HSP90 (diluted 1:20,000, Proteintech, Catalog: 60318-1-Ig), β-actin (diluted 1:50,000, Proteintech, Catalog: 66009-1-Ig), HBB, MMP13 (diluted 1:2000, Proteintech, Catalog: 18165-1-AP), SOD2 (diluted 1:20,000, Proteintech, Catalog: 24127-1-AP) and Collagen Type II.

Techniques: Knockdown, Western Blot, Analysis, Transfection, Over Expression, Plasmid Preparation, Staining, Fluorescence