c28 i2 cells  (TargetMol)

Journal: Frontiers in Pharmacology

Article Title: Pterostilbene attenuates osteoarthritis progression through p53-dependent autophagy activation: evidence from network analysis and experimental validation

doi: 10.3389/fphar.2026.1686555

Figure Lengend Snippet: PT modulates p53 localization, AMPK/mTOR signaling, and autophagy-related markers in chondrocytes. (A) CETSA results assessing the thermal stability of p53 in C28/I2 cells treated with PT or vehicle across a temperature range of 38 °C–70 °C. Representative WB bands are shown above the normalized thermal aggregation curves; (B) Representative immunofluorescence confocal microscopy images illustrating p53 subcellular localization (red) and DAPI-stained nuclei (blue) in chondrocytes across different treatment groups (scale bar = 10 μm); (C) Representative WB bands showing p53 distribution (nuclear, cytoplasmic, and total), the phosphorylation status of AMPK and mTOR, and autophagy markers (LC3 and p62); (D–K) Quantitative analyses of protein expression levels: total p53/GAPDH (D) , nuclear p53/PCNA (E) , cytoplasmic p53/GAPDH (F) , nuclear/cytoplasmic p53 ratio (G) , p-AMPK/AMPK (H) , p-mTOR/mTOR (I) , p62/GAPDH (J) , and the LC3II/I ratio (K) ; (L) Representative TUNEL staining images showing apoptotic cells (red) and nuclei (blue) (scale bar = 50 μm); (M) Quantitative analysis of the percentage of TUNEL-positive cells across groups. Data = mean ± SEM (n = 3). **P < 0.01, ***P < 0.001 vs. control; ###P < 0.001 vs. IL-1β; †P < 0.05, ††P < 0.01, †††P < 0.001 vs. IL-1β + PT.

Article Snippet: C28/I2 cells were treated with PT (HY-N0828, Lot# 130513, MedChemExpress, Shanghai, China) without further purification.

Techniques: Immunofluorescence, Confocal Microscopy, Staining, Phospho-proteomics, Expressing, TUNEL Assay, Control

Journal: Journal of Advanced Research

Article Title: Panaxatriol exerts anti-senescence effects and alleviates osteoarthritis and cartilage repair fibrosis by targeting UFL1

doi: 10.1016/j.jare.2024.10.016

Figure Lengend Snippet: UFL1 is associated with celluar senescence and Panaxatriol modulates chondrocyte senescence through UFL1 . (A) Western blot was used to detect the protein level of UFL1, P21 and SASPs (MMP9,MMP13,ADAMTS-4,IL-6,TNF-α) in UFL1-KO cells. (B) Representative images of immunofluorescence staining of P21 and γ-H2AX of knee joint sections of DMM mice treated with DMSO or panaxatriol and the quantification results. (C) Representative images of immunofluorescence staining of P21 and γ-H2AX in the sections of human cartilage explants treated with 10 ng/mL IL-1β or a series concentration of panaxatriol for 7 days. (D) Representative images of SA-beta-gal staining C28/I2 cells treated with 10 ng/mL IL-1β or a series concentration of panaxatriol for 2 days. (E) Based on the UFL1 knock-out C28I2 cells (UFL1-KO), western blot was used to detect the influence of UFL1 on the ability of panaxatriol to regulate P21 and SASPs. OE-UFL1 means overexpression of UFL1 in UFL1-KO cells. (F) Based on UFL1-KO cells, western blot was used to detect the influence of UFL1 on the ability of panaxatriol to regulate P65,p-P65 and FOXO1.The values are mean ± SEM of at least three independent experiments. (*,P < 0.05; **,P < 0.01;***,P < 0.001; ****,P < 0.0001;ns, no significance).

Article Snippet: For inhibitor and agonist treatment, C28/I2 cells were treated with or without 10 μM JSH-23 (TargetMol, Cat.T1930), 1 μM Phorbol 12-myristate 13-acetate (PMA,TargetMol, Cat.TQ0198), 10 μM AS1842856 (MedChemExpress, Cat.HY-100596).

Techniques: Western Blot, Immunofluorescence, Staining, Concentration Assay, Knock-Out, Over Expression