Fig. 3: circPKNOX1 acted as a sponge for miR-370-3p in NP cells.
From: Inhibition of intervertebral disc disease progression via the circPKNOX1–miR-370-3p–KIAA0355 axis
A Through the intersection of TargetScan, miRanda, and RNA hybrid data, circPKNOX1 target microRNA was predicted. B After knocking down circPKNOX1, the expression level of hsa-miR-370-3p increased; *P<0.05. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test. C The expression level of miR-370-3p increased in degenerated tissues (six different donors); *P<0.05. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test. D Left: the fluorescence intensity of miR-370-3p increased in degenerated tissues, as determined using fluorescence in situ hybridization (scale bar, 25–100μm). Right: the fluorescence intensity was quantitatively analyzed (two different donors); ***P<0.01. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test. The miR-370-3p probe was labeled with Alexa Fluor 488. E miR-370-3p was located in the cytoplasm and intersected with circPKNOX1 in the cell substructure (scale bar, 25μm). F The binding site of circPKNOX1 and miR-370-3p was identified using CircInteractome. G The efficiency of miR-370-3p mimic and inhibitor was detected; *P<0.05. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test. H The expression of related proteins (collagen II, aggrecan, SOX9, MMP3, MMP13, ADAMTS4, and ADAMTS-5) was detected using western blotting. I The fluorescence intensity of related proteins (collagen II, aggrecan, ADAMTS4, and MMP13) was detected via an immunofluorescence assay (scale bar, 100μm). J RT-qPCR was used to detect the expression of related genes; *P<0.05, **P<0.01, ***P<0.001. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test. K Left: the extracellular matrix was detected by Alcian staining. Right: the Alcian staining was quantitatively analyzed. Data represent mean±SD, and the P values were determined by a two-tailed unpaired Student’s t test.
