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The Predicted Target Genes of miR-122/449a Validation and Effects on Sertoli Cells Proliferation

The Predicted Target Genes of miR-122/449a Validation and Effects on Sertoli Cells Proliferation

Lihong Qin1, Guoliang Zhang1, Chaojie Zhu1, Jian Wu1, Zhihui Zhao2,* and Yumin Zhao1,*

1Jilin Academy of Agricultural Sciences, Xixinghua Road 186, Gongzhuling 136100, China
2Agricultural College, Guangdong Ocean University, Zhanjiang 524088, China

*      Corresponding authors: a840581260@126.com;
yumin_zh@163.com

 

Fig. 1.

Target sites of miR-122/499a prediction and luciferase assay. A, miR-122 and miR-449a target genes prediction results; B, the target relationship between miR-122 and DUSP4/PDK4, miR-449a and FKBP1B detected by dual-luciferase reporter system. NC, blank vector; WT, wild-type vector; mut, mutant vector (**p<0.01).

Fig. 2.

The genes expression levels in different months of testicular tissues. A, relative expressions of miR-122 and miR-449a in 1-day, 12-months-old and 24-months-old bull testes tissues; B, the mRNAs expressions of PDK4, DUSP4 and FKBP1B in 1-day, 12-months-old and 24-months-old bull testes tissues (**p<0.01).

Fig. 3.

miR-122/449a and their target genes expression levels detection in bull testicular Sertoli cells. A, miR-122 and DUSP4, PDK4 expression levels were detected in Sertoli cells that transfected with miR-122 mimics, miR-122 inhibitor and miR-shNC (**p<0.01); B, miR-449a and target gene FKBP1B expression levels were detected in cells that transfected miR-449a mimics, miR-449a inhibitor and miR-shNC (**p<0.01); C, Western blot analysis for DUSP4, PDK4 and FKBP1B after miR-122 and miR-449a transfection.

Fig. 4.

The Sertoli cells proliferation was inhibited by DUSP4, PDK4 or FKBP1B. The DUSP4, PDK4 or FKBP1B recombinant plasmid was transfected into Sertoli cells. A, the mRNA expression levels of DUSP4, PDK4 and FKBP1B; B, Western blot detection results for DUSP4, PDK4 and FKBP1B; C, MTT assay results after recombinant plasmid overexpression (**p<0.01).

Fig. 5.

The Sertoli cells proliferation was promoted via miR-122/PDK4/DUSP4 and miR-449a/FKBP1B pathways. A, Sertoli cells proliferation was analyzed by MTT assay after the cells were transfected with miR-122 mimics, miR-122 inhibitor and miR-shNC; B, Sertoli cells proliferation was analyzed by MTT assay after the cells were transfected with miR-449a mimics, miR-449a inhibitor and miR-shNC (**p<0.05, **p<0.01).

Pakistan Journal of Zoology

June

Vol. 50, Iss. 3, Pages 799-1198

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