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Microarray and Bioinformatics Analysis of Differentially-Expressed MicroRNAs in Wnt/Β-Catenin Signaling-Activated Bovine Skeletal Muscle Satellite Cells

Microarray and Bioinformatics Analysis of Differentially-Expressed MicroRNAs in Wnt/Β-Catenin Signaling-Activated Bovine Skeletal Muscle Satellite Cells

Ming-Yan Shi2, Xu-Liang Yan1, Ping-Ping Han1, Yu-Jie Tang1 and Hui-Xia Li1*

1College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China 
2College of Life Science, Luoyang Normal University, Luoyang, 471934, China
 
Xu-Liang Yan and Ming-Yan Shi are co-first authors.

*      Corresponding author: lihuixia@njau.edu.cn

Fig. 1.

(A) Bovine skeletal muscle satellite cells (bSMSCs) cultured in vitro; primitive bSMSCs were rounded and had strong refraction; (B) The fast-adhered cells were fibroblasts and the bSMSCs were round and suspended; (C) Two days later, the bSMSCs were spindles and proliferating rapidly; (D, E, F) After two cell passages and differential adhesion, the cells were confluent and became homogeneous in their appearance.

Fig. 2.

Immunofluorescence staining showed the expression of pax7. (A, D) Staining nuclei with Hoechst; (B, E) Using anti pax7 antibody immunofluorescence staining, 95% of the cells were positive; (C, F) Pax7 and Hoechst composite picture. (A, B, C bar = 10 μm; D, E, F bar = 50 μm).

Fig. 3.

Skeletal muscle satellite cells were treated with LiCl to activate Wnt signaling. (a) Wnt10b and (b) β-catenin microRNA expression levels were detected by quantitative real-time PCR. * P < 0.05; **P < 0.01.

Fig. 4.

The spot falling on graph y=x line (the median line on the graph) represents the probe point signal values differences in the two chips (fold change=1). Graph points fall on both sides of the bit line, which is outside the 45° line, representing the difference between the value of the probe signal point in the two chips (fold change >2). An area: 2x increased microRNAs (miRNAs); B area: 2x decreased miRNAs.

Fig. 5.

Expression of miRNAs by quantitative real-time PCR (qRT-PCR) and microarray signal intensity. (a) Relative detectable signal intensity of differentially expressed miRNAs between different models using microarray. (b) Relative expression levels of differentially expressed miRNAs by qRT-PCR. *P < 0.05; **P < 0.01.

Pakistan Journal of Zoology

December

Vol. 52, Iss. 6, Pages 2027-2426

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