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. 2021 Jan 13;11(1):e255. doi: 10.1002/ctm2.255

FIGURE 5.

FIGURE 5

FoxO3 was a direct target of miR‐29b‐3p. A, Binding sequences of miR‐29b‐3p and FoxO3 were predicted by bioinformatics analysis. B, Binding of miR‐29b‐3p and FoxO3 was confirmed by the dual‐luciferase reporter assay. *P < .05 compared with the miR‐NC group. C, miR‐29b‐3p overexpression reduced FoxO3 level, while miR‐29b‐3p inhibition increased FoxO3 level. D, FoxO3 overexpression increased the protein level of Col II, aggrecan, and Sox9 and decreased the level of MMP13, ADAMTS5, and Runx2. E, Chondrocytes were treated with miR‐29b mimics with or without overexpressing FoxO3 and stimulated by IL‐1β (10 ng/mL) challenge for 24 hours. Apoptosis was assessed by flow cytometry. F and G, Light microscopy images and quantitative analysis of chondrocytes stained with the senescent marker SA‐β‐Gal in monolayer culture; scale bar: 100 μm. H, Light microscopy images and number of transmitted cells in the transwell migration assay. I and J, Light microscopy images and quantitative analysis of scratch wound assays. K and L, Protein and mRNA level of genes associated with chondrocytes (MMP13, ADAMTS5, Runx2, Col II, aggrecan, and Sox9). M, Immunofluorescence for MMP13, Col II, and aggrecan. The data are shown as mean ± SD (n = 3; *P < .05, **P < .01)