Figure 8. (A) Bioinformatic prediction of binding site of miR-181a on DLEU2. (B, C) Protein expression of SEPP1 in C2C12 cells transfected with miR-181a inhibitor or miR-181a mimic as determined by Western blot assay. * P < 0.05 vs control, each test was performed in triplicate. (D) Gene expression of SEPP1, MyoD and MyoG in C2C12 cells transfected with miR-181a inhibitor or miR-181a mimic as determined by RT-PCR assay. *P < 0.05 vs control, each test was performed in triplicate. (E) Analysis of miR-181a regulation by SEPP1 using Luciferase reporter assays. *P < 0.05 vs miR-NC. (F) Effect of DLEU2 overexpression, miR-181a inhibitor and miR-181a mimic on the proliferation of C2C12 cells. (G) C2C12 myoblasts were treated with miR-181mimic or miR-181inhibitor. Cells were stained with Edu. Quantification of relative ratio of Edu+ C2C12 cells. Data are presented as the mean ± S.D. (n = 3). ***p<0.005, ****p<0.0005. (H) A model showing the inhibitory effect of lncDLEU2-miR-181a-SEPP1 pathway in muscle differentiation and proliferation. LncRNA DLEU2 as a miR-181a sponge regulates SEPP1 expression and inhibits muscle differentiation and regeneration.