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. 2022 Sep 16;7:53. doi: 10.1038/s41536-022-00250-7

Fig. 4. SOX11 is a direct target of miR-455 to modulate chondrogenesis.

Fig. 4

A All predicted genes were compiled for Venn analysis to search for the potential targets of miR-455. B miRNA–mRNA network using the Cytoscape software was constructed for miRNA-455. C Sequence of wild-type (WT) and mutant (Mut) SOX11 binding sites for miRNA-455 (left) and conservation level of miR-455 sequence among species (right). D, E SOX11 expression(red) assayed with fluorescent immunostaining (red for SOX11; green for cytoskeleton) and qRT-PCR in T3-EV treated BMSC recipient (n = 3 for each with the same BMSCs and the same EVs). F Fluorescence micrograph of Cy3 (red)-labeled miR-455 mimic internalized by BMSCs. G Luciferase reporter assay analysis results (n = 3 for each) to confirm the direct interaction between miR-455 and SOX11 binding sites. Relative luciferase reporter activity was assessed for co-transfected SOX11 WT (or Mut) with miR-455 mimics or inhibitors in cultured primary BMSC cells. miRNA-455 mimics control and inhibitor control served as negative controls. H–J miR-455 and SOX11 expression(red) with H, J) qRT-PCR and I fluorescent immunostaining in BMSCs transfected with miR-455 mimics or inhibitor (n = 3 for each). K, L Fluorescent immunostaining was also conducted on anabolic and catabolic markers ACAN (green) and MMP13 (green) in BMSCs transfected with miR-455 mimics or inhibitors. Cells were counterstained with DAPI for the nucleus (blue). *P < 0.05, **P < 0.01, ***P < 0.001, NS not significant.