SOX9 Antibody - #AF6330

FIGURE 5 Pathogenic FLS EVPs impair chondrogenic differentiation of mesenchymal stem cells. (A) Schematic diagram of EVP stimulation on mouse BMSCs isolated from the femoral bone marrow cavity of mice. (B) Internalization of EVPs by BMSCs. Scale bar = 50 µm. (C) Expression of chondrogenic differentiation-related genes in chondrogenesis-induced BMSCs after 7 days of treatment with different EVPs. (D) Representative Western blot images of COL10A1, the marker of chondrocyte hypertrophy. (E, F) Representative images of alcian blue staining and SA-β-gal staining in BMSCs after 7 days of stimulation with different EVPs. Scale bar = 1 mm and 200 µm separately. (G) Schematic diagram of section staining observation after inducing BMSCs to form chondrocyte pellets for 21 days while simultaneously stimulating with different EVPs. (H, I) Representative images of SOX9 fluorescence staining, safranin O (SO) staining, alcian blue (AB) staining and toluidine blue (TB) staining of BMSC-differentiated chondrocyte pellet sections. Scale bar = 50 µm. (J) Schematic diagram of EVP treatment on mouse ADSCs isolated from the iWAT of mice. (K) Internalization of EVPs by ADSCs. Scale bar = 50 µm. (L, M) Representative images of SA-β-gal staining and alcian blue staining of ADSCs after 7 days of chondrogenic induction and treatment with different EVPs. (N) Representative images of SOX9 staining in sections of chondrocyte pellets formed by ADSC after 21-day induction.

Fig. 7 The chondrogenic differentiation of iPSCs with different Li+ ions concentration in 14 days. a Immunofluorescence of chondrocytes specified proteins (COL II, Aggrecan, and SOX9) and hypertrophic specified protein (COL X and MMP13). b The average fluorescence intensity of COL II, Aggrecan, SOX9, MMP13, and COL X proteins in each group, n = 5. c The relative genes expression of chondrogenic differentiation cultured with Li+ ions, n = 3. d The relative genes expression of hypertrophic differentiation cultured with Li+ ions, n = 3. Data presented as mean ± SEM. Scale bar = 200 μm. CTR: MCDM without any Li+ ions. *p < 0.05, **p < 0.01, ***p < 0.001

Fig. 8. GPX4 inhibitor RSL3 substantially attenuates VK2 protection of chondrocytes. (A-I) WB results for GPX4, NFκB, pMAPK/MAPK, Aggrecan, CollagenⅡ, SOX9, MMP3, MMP13 and Tubulin; (J) Quantitive analysis of relative Glutathione Peroxidase Activity; (K-L) GSH contents and ratio of GSH/GSSG; (M) Quantitative results of MDA content assay.

Figure 3. Key indicators expression of knee cartilage in 7-day male mice offspring of control and obese groups. (A) Relative mRNA, (B) representative western blot bands, and (C) statistical results of knee cartilage 7-day male mice offspring in the control group versus obese group. O-WT-P7: offspring 7-day male mice from the control mothers. O-OB-P7: offspring 7-day male mice from the obese mothers. (A) n = 6 represents six mice in each group from at least three litters of different mothers. (B) n = 6, each band represents the expression of cartilage tissue proteins in two mice from at least three litters of different mothers. Results are expressed as mean ± SD. ns not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001.