MicroRNA-181a Regulates the Proliferation and Differentiation of Hu Sheep Skeletal Muscle Satellite Cells and Targets the YAP1 Gene

Figures

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  Figure 1. MiR-181a suppresses SMSCs’ proliferation in Hu sheep: (a,b) The relative expression of miR-181a; (c,d) The relative mRNA expression of PCNA and CDK2; (e,f) CCK-8 assay of SMSCs; (g,h) Cell-cycle analysis of SMSCs; (i,j) The relative protein expression of CDK2; (k,l) Statistical analysis of (i,j); (m,n) EdU assay of SMSCs. Fluorescence inverted microscope (Nikon, Tokyo, Japan) was used to obtain microscopic images. The magnification is 100 times; (o,p) Statistical analysis of EDU-stained cells in microscopic images of (m,n). In all graphs, all data are presented as means ± SEM (standard error of the mean) (n = 3). Statistical significance was performed by unpaired Student’s t-test. (* p < 0.05; ** p < 0.01; ns p > 0.05) vs. NC (negative control).

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  Figure 2. MiR-181a facilitates SMSCs differentiation in Hu sheep: (a) Differentiation images of SMSCs in vitro; (b) The relative expression of miR-181a during SMSCs’ differentiation (DM0-DM4 represents differentiation from zero to four days); (c,d) The relative mRNA expression of MyHC, MyoD, and MyoG; (e,f) The relative protein expression of MyHC; (g,h) Statistical analysis of (e,f); (i,j) Immunofluorescence staining of SMSCs. The magnification is 100 times. In all graphs, all data are presented as means ± SEM (standard error of the mean) (n = 3). Statistical significance was performed by unpaired Student’s t-test. (* p < 0.05; ** p < 0.01) vs. NC (negative control).

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  Figure 3. YAP1 is a Target Gene of miR-181a: (a) Homology analysis of miR-181a in different species; (b) RNAhybrid predicted the targeted binding relationship between YAP1 and miR-181a; (c) The interaction model between miR-181a and YAP1 3′UTR by RNAhybrid online software; (d) The luciferase assays of HEK293T; (e,f) The mRNA expression of YAP1; (g,h) The protein expression of YAP1; (i,j) Statistical analysis of (g,h). In all graphs, all data are presented as means ± SEM (standard error of the mean) (n = 3). Statistical significance was performed by unpaired Student’s t-test. (* p < 0.05; ** p < 0.01; *** p < 0.01; ns p > 0.05) vs. NC (negative control).