miR-31 from Mesenchymal Stem Cell-Derived Extracellular Vesicles Alleviates Intervertebral Disc Degeneration by Inhibiting NFAT5 and Upregulating the Wnt/β-Catenin Pathway

Figure 6

miR-31 targets NFAT5 and activates the Wnt/β-catenin pathway in NPCs: (a) a PPI network of the genes related to the key downstream genes constructed through GeneMANIA; (b) binding of miR-31 to NFAT5 in 293T cells determined by dual luciferase reporter assay; (c) expression of miR-31 in NPCs after transfection with miR-31 mimic or miR-31 inhibitor determined by RT-qPCR; (d) expression of NFAT5 in NPCs after transfection with miR-31 mimic or miR-31 inhibitor determined by RT-qPCR; (e) expression of NFAT5 and β-catenin in NPCs after transfection with miR-31 mimic or miR-31 inhibitor determined by Western blot; (f) the transcription activity of TCF/LEF in NPCs after transfection with miR-31 mimic or miR-31 inhibitor determined by TOPFlash; (g) protein expression of NFAT5 and β-catenin in TNF-α-treated NPCs determined by Western blot; (h) the transcription activity of TCF/LEF in TNF-α-treated NPCs determined by TOPFlash; (i) expression of NFAT5 and β-catenin in TNF-α-treated NPCs cocultured with BMSCs-EVs or BMSCs-EVs+anti-miR-31 determined by Western blot; (j) the transcription activity of TCF/LEF in TNF-α-treated NPCs cocultured with BMSCs-EVs or BMSCs-EVs+anti-miR-31 determined by TOPFlash. . Cell experiments were repeated three times.

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