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. 2022 Apr 12;2022:3156245. doi: 10.1155/2022/3156245

Figure 1.

Figure 1

Kindlin-2 promotes chondrogenesis in the direct contact coculture system. (a) A direct contact BMSC-chondrocyte coculture system to evaluate the effects of BMSCs on chondrocytes. (b) Heatmap of DEGs of chondrocytes cocultured with or without BMSCs. Green and red colors represent low and high expression values, respectively. (c) Representative biological process (BP) categories identified in GO analyses based on upregulated DEGs in cocultured chondrocytes compared with control counterparts. (d) Expression pattern of Kindlin-2 in chondrocytes cocultured with or without BMSCs were determined using RT–qPCR. Values are expressed as mean ± s.d., ∗∗∗p < 0.001. (e) The knockdown efficiency of Kindlin-2 in chondrocytes was confirmed by qPCR and Western blotting. Values are expressed as mean ± s.d., ∗∗p < 0.01, ∗∗∗p < 0.001, ns indicates no significance. (f) Overexpression efficiency of Kindlin-2 in chondrocytes was confirmed by qPCR and Western blotting. Values are expressed as mean ± s.d., ∗∗∗p < 0.001. (g) mRNA expression levels of cartilage-specific genes (Col2, Sox-9, and Aggrecan) in chondrocytes on day 7 were detected by qPCR in different groups. 18S was used as an internal control. Values are expressed as mean ± s.d., ∗∗p < 0.01, ns indicates no significance. (h) In a direct contact coculture system for 7 days, knockdown of Kindlin-2 attenuated the regulation of BMSCs on chondrocyte ECM secretion, as indicated by Alcian blue staining and safranin O staining. Chondrocytes without coculture were used as the control (Con) group. (i, j) Quantitative evaluation of Alcian blue staining results (i) and safranin O staining results (j) on day 7. Values are expressed as mean ± s.d., ∗∗p < 0.01, ∗∗∗p < 0.001, ns indicates no significance. (k) mRNA expression levels of Col2, Sox-9, and Aggrecan in chondrocytes on day 7 by qPCR in different groups. 18 s was used as an internal control. Values are expressed as mean ± s.d., ∗∗∗p < 0.001, ns indicates no significance. (l) In the direct contact coculture system, Kindlin-2-overexpressing chondrocytes enhanced chondrogenic ability on day 7, as revealed by Alcian staining and safranin O staining. Chondrocytes cultured alone were set as the Con group. Vec: vector group; OE: overexpression group. (m, n) Quantitative analyses of Alcian staining results (m) and safranin O staining results (n) on day 7 were performed. Values are expressed as mean ± s.d., ∗∗∗p < 0.001.