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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: FTO-mediated m6A demethylation of pri-miR-3591 alleviates osteoarthritis progression

Fig. 4

miR-3591-5p aggravated LPS-induced chondrocytes damage in vitro by targeting regulation of PRKAA2. A Common target genes predicted by TargetScan and miRBD were intersected with the significantly downregulated mRNA in OA cartilage from GSE113825 and GSE16464 analysis. B The expression levels of miR-3591-5p and PRKAA2 were tested by RT-qPCR in chondrocytes with miR-3591-5p mimics or inhibitor. C RIP-Ago2 analysis indicated that interaction between miR-3591-5p and PRKAA2. D Interaction between miR-3591-5p and PRKAA2 was affirmed by the dual-luciferase reporter assay. E Chondrocytes were transfected with miR-3951 inhibitor or NC inhibitor, and then treated with 40 ng/mL LPS for 24 h. RT-qPCR was used to detect the mRNA expression miR-3591-5p and PRKAA2. F Chondrocytes were transfected with miR-3951 inhibitor or a combination of miR-3951 inhibitor and shPRKAA2 and then treated with 40 ng/mL LPS for 24 h. CCK-8 was used to detect the viability of chondrocytes. G Chondrocytes were transfected with miR-3951 inhibitor or a combination of miR-3951 inhibitor and shPRKAA2 and then treated with 40 ng/mL LPS for 24 h. The apoptosis of chondrocytes was tested by flow cytometry. H Chondrocytes were transfected with miR-3951 inhibitor or a combination of miR-3951 inhibitor and shPRKAA2 and then treated with 40 ng/mL LPS for 24 h. Western blotting was applied to assess the expression of MMP13, ADAMTS5, Aggrecan, and COL2A1. LPS, lipopolysaccharide; NC mim., negative control corresponding to miR-3591-5p mimics adenovirus; miR mim., miR-3591-5p mimics adenovirus; NC inh., negative control corresponding to miR-3591-5p inhibitor adenovirus; miR inh., miR-3591-5p inhibitor adenovirus; shPRKAA2, PRKAA2 knockdown adenovirus; n = 3, *p < 0.05, **p < 0.01, ***p < 0.001

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