WTAP-mediated N6-methyladenosine modification of circ_PVT1 promotes proliferation and tenogenic differentiation of tendon stem/progenitor cells

Figure 2 Knockdown of circular RNA plasmacytoma variant translocation 1 reversed the transforming growth factor-β1-induced tenogenic differentiation in tendon stem/progenitor cells. A and B: Western blot analysis of scleraxis, tenomodulin, and COL3; C: Cell Counting Kit-8 assay for the detection of cell proliferation in tendon stem/progenitor cells (TSPCs) of the negative control (NC), Transforming growth factor (TGF)-β1, TGF-β1 + shNC, and TGF-β1 + circular RNA plasmacytoma variant translocation 1 (circ_PVT1) groups; D and E: Transwell assay for migration analysis of TSPCs in the NC, TGF-β1, TGF-β1 + shNC, and TGF-β1 + circ_PVT1 groups. Scale bars = 200 μm; F and G: Detection and analysis of apoptosis in TSPCs of the NC, TGF-β1, TGF-β1 + shNC, and TGF-β1 + circ_PVT1 groups; H: TGF-β1 treatment promoted the expression of circ_PVT1 in TSPCs. After treatment with 5 ng/mL of TGF-β1 for 1 day, 3 days, or 7 days, the expression of circ_PVT1 was increased compared with that of the normal control; I: Knockdown efficiency of shNC and various shRNAs of circ_PVT1 as determined by quantitative polymerase chain reaction in TGF-β1-induced TSPCs; J and K: The Picro-Sirius red staining of collagen in TSPCs of the NC, TGF-β1, TGF-β1 + shNC, and TGF-β1 + circ_PVT1 groups. Scale bars = 200 μm. All data are presented as the mean ± SD. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. SCX: Scleraxis; TNMD: Tenomodulin; NS: No significance; NC: Negative control; TGF-β1: Transforming growth factor-β1; circ_PVT1: Circular RNA plasmacytoma variant translocation 1.