Original Image of Guzhi Zengsheng Zhitong Decoction Mitigates Osteoarthritis via FOXO1-Mediated Autophagy and Apoptosis
Description
figure 1 (B-D) Representative histological images of knee joints after 4 weeks of treating, including hematoxylin-eosin (HE), safranin-O fast green, and toluidine blue staining. figure 2 (A) Micro-CT images showing 3D reconstructions of the distal femur and transverse and sagittal views of the tibial plateau. figure 6 (I, J) Immunohistochemical analysis and quantification of FoxO1, LC3B, Parkin and PINK1 expression in the rat articular cartilage following oral administration (n=4, scale bar: 200 μm). figure 7 (A) Representative transmission electron microscopy images showing mitochondrial morphology (scale bars: 5 μm, 2 μm, 500 nm). (C) Fluorescence images of mitochondrial membrane potential (MMP) staining. Red fluorescence represents JC-1 aggregates, green fluorescence represents JC-1 monomers, and the red/green intensity ratio reflects MMP (scale bar: 100 μm). (D) Fluorescence images of mitochondrial permeability transition pore (MPTP) staining (scale bar: 200 μm). (E) Fluorescence images of reactive oxygen species (ROS) levels (scale bar: 200 μm). figure 8 (C) TUNEL staining to evaluate apoptosis in knee cartilage of OA rats after oral GZZD treatment. (scale bar: 100 μm). figure 10 (A) JC-1 staining showing mitochondrial membrane potential (MMP) in OA chondrocytes with and without FoxO1 silencing (scale bar: 200 μm). (C) Effects of FoxO1 silencing on mitochondrial MPTP permeability after GZZD treatment. (scale bar: 200 μm). (E) ROS levels visualized by fluorescence staining before and after FoxO1 silencing. (scale bar: 200 μm). Supplement figures Figure S1. Representative anatomical images of rat knee joints at week 4 post-treatment. Figure S2. Identification of primary chondrocytes. (A) Observation of cell morphology; (B) COL2A1 immunofluorescence staining. Figure S5. Effects of GZZD on OA chondrocyte apoptosis observed via Annexin V-FITC/PI staining (in vitro). Figure S8. Effects of oral GZZD on STAT3, FOXO1, P-P65, and P65 protein levels in OA rat chondrocytes analyzed by Western blot and quantitative analysis, in vivo Western Blot Original Image.