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. 2023 Jan 5;13(1):e1158. doi: 10.1002/ctm2.1158

FIGURE 3.

FIGURE 3

CircZSWIM6 knock‐down or overexpression governs extracellular matrix metabolism and senescent phenotype: (A) human chondrocyte transfected with three CircZSWIM6 small interfering RNA (siRNA); knock‐down efficiency indicated by quantitative real‐time polymerase chain reaction (RT‐qPCR) (n = 3, three donors for three replicates); no change of ZSWIM6 mRNA expression is detected; (B and C) mRNA level of Col2, Sox9, MMP13, ADAMTS5, p16, p21 and p53 in human chondrocyte with CircZSWIM6 knock‐down (n = 3, three donors for three replicates); (D) protein level of Col2, Sox9, p21, p16, p53, MMP3 and ADAMTS5 in human chondrocyte after CircZSWIM6 knock‐down; (E) senescence β‐galactosidase (SA‐β‐Gal) staining, immunofluorescence images of aggrecan, p16 and MMP13 in chondrocytes after CircZSWIM6 knock‐down. (F) Alcian blue and Toluidine blue staining in human chondrocyte with CircZSWIM6 knock‐down, (G) SA‐β‐Gal of human chondrocyte with CircZSWIM6 knock‐down or doxorubicin stimulated human chondrocyte with or without CircZSWIM6 knock‐down: quantification of SA‐β‐Gal, aggrecan, p16 and MMP13 stainings (n = 6); (H) CircZSWIM6 overexpression efficiency (n = 3, three donors for three replicates); (I) protein level of Col2, Sox9, p21, p16, p53, MMP13 and ADAMTS5 in human chondrocyte with CircZSWIM6 overexpression; (J) SA‐β‐Gal staining in chondrocyte after CircZSWIM6 overexpression; (K) Alcian blue staining in chondrocyte after CircZSWIM6 overexpression; (L) mRNA level of Col2, Sox9, p21, p16, p53, MMP13 and ADAMTS5 in human chondrocyte with CircZSWIM6 overexpression; (M and N) quantifications of SA‐β‐Gal and Alcian blue staining after CircZSWIM6 overexpression; quantitative data shown as mean ± SD; exact p‐values are shown in figures. Two‐side unpaired Student's t‐test was used for statistical analysis. One‐way ANOVA with Tukey's multiple comparison was used for statistical analysis.