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miR-216b Downregulates Intestinal IL2RB to Inhibit the Invasion of Echinococcus granulosus Eggs in the Kazakh Sheep

miR-216b Downregulates Intestinal IL2RB to Inhibit the Invasion of Echinococcus granulosus Eggs in the Kazakh Sheep

Xuhai Wang1,2, Xin Li1, Fangyuan Yuan1, Chaocheng Li1, Bin Jia1* and Song Jiang1*

1College of Animal Science and Technology, Shihezi University, Road Beisi, Shihezi 832003, Xinjiang, P. R. China.
2Chongqing Mintai New Agrotech Development Group Co., Ltd. Mintai road, shilong industrial park, jiguanshi, Nanan district 400063, Chongqing. P. R. China.

*      Corresponding author: jiabin@shzu.edu.cn; jiangsong918@shzu.edu.cn

Fig. 1.

The distribution of IL2RB in the small intestine of sheep. Intestinal IL2RB expression in NRG and RG sheep was determined by immunohistochemical staining. A, C: Strong IL2RB positive expression was observed in T cells of mucosal layer and musclar layer. B, D: The positive signal of IL2RB was only detected in mucosal layer, especially in the lamina propria and epithelium. E: The average IOD was obtained by analysing IL2RB IHC in four random felds of each slide. Upper panel: representative image; lower panel: quantitative analysis. Results are presented as the mean± SEM (magnifcation, 200×, scale bars= 100μm).

Fig. 2.

miR-216b and IL2RB protein levels are inversely correlated in RG and NRG sheep intestinal tissues. miRNA in situ hybridization for miR-216b was performed on intestinal sections from RG and NRG controls (green, miR-216b; blue, DAPI nuclear staining). (Magnifcation 200×, scale bar= 200μm).

Fig. 3.

The IL2RB 3’-UTR contains conserved miR-216b target sites. A: The fold-changes of seven miRNAs predicted by three algorithms in RG intestinal tissues compared to NRG (control), as analysed by high-throughput sequencing. B: Schematic illustration of conserved duplexes formed by IL2RB 3’UTR and miR-216b interactions. The predicted free energy of the hybrid is noted. Paired bases are marked by a blue line. The complementary seed sites are marked in blue. C: Quantitative real-time PCR analysis of IL2RB expression in 293T cells transfected with miR-216b mimics, miR-216b inhibitor, IL2RB-W or IL2RB-M. Data are presented as the mean±SEM from three independent experiments (293T group: ***p=0.000092, IL2RB-M vs. IL2RB-W). D: Relative luminescence intensity detected by a Hamamatsu optical analyze reader after miR-216b mimics and dual-luciferase vectors were co-transfected into 293T cells (*** P<0.01).

Pakistan Journal of Zoology

October

Vol. 53, Iss. 5, Pages 1603-2000

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