FIGURE SUMMARY
Title

miR-19b-3p regulated by estrogen controls lipid synthesis through targeting MSMO1 and ELOVL5 in LMH cells

Authors
Jia, Q., Cao, Y., Zhang, M., Xing, Y., Xia, T., Guo, Y., Yue, Y., Li, X., Liu, X., Zhang, Y., Li, D., Li, Z., Tian, Y., Kang, X., Li, H.
Source
Full text @ Poult. Sci.
Fig. 1

Conservative analysis and spatiotemporal expression profile of miR-19b-3p. (A) Alignment of miR-19b-3p mature sequence among species. (B) The relative expression of miR-19b-3p in different tissues. (C) The relative expression of miR-19b-3p in liver of chickens at different stages. U6 was used as an internal reference gene to estimate the relative expression of miRNA (n = 6). Different lowercase letters in each bar denote a significant difference (P < 0.05).

Fig. 2

Effects of overexpression miR-19b-3p on lipid related indicators and genes in LMH. (A) The overexpression efficiency of miR-19b-3p mimics. (B) oil red o staining of LMH cells treated with miR-19b-3p mimics or mimics NC. (C) Semiquantitative assessment of oil red o content absorbance detection at 510 nm. (D) Effects of overexpression miR-19b-3p on TG and TC contents in LMH cells. (E) Effects of overexpression miR-19b-3p on the expression of lipid metabolism-related genes. Mimics NC means the negative control of the miR-19b-3p mimics; TG indicates triglycerides, TC indicates cholesterol; * means P < 0.05, ** means P < 0.01.

Fig. 3

Effects of interference miR-19b-3p on lipid related indicators and genes in LMH. (A) The interference efficiency of miR-19b-3p inhibitor. (B) Oil Red O staining of LMH cells treated with miR-19b-3p inhibitor or inhibitor NC (C). Semiquantitative assessment of Oil Red O content absorbance detection at 510 nm. (D) Effects of interference miR-19b-3p on TG and TC contents in LMH cells. (E) Effects of interference miR-19b-3p on the expression of lipid metabolism-related genes. TG indicates triglycerides and TC indicates cholesterol

Fig. 4

Prediction and screening of miR-19b-3p target genes. (A) Prediction analysis of miR-19b-3p target genes. (B) Screening of potential candidate genes by overexpression or interference miR-19b-3p. (C) Expressions of miR-19b-3p, MSMO1 and ELOVL5 in liver of chickens at different stages.

Fig. 5

Conservative analysis of miR-19-3p binding sites and ELOVL5/MSMO1 motifs among species. (A) Prediction of minimum binding free energy between miR-19b-3p and potential target gene ELOVL5/MSMO1. (B) Conservative analysis of miR-19b-3p binding sites in MSMO1/ELOVL5 3′UTRs among different species. (C) Conservative motifs analysis of ELOVL5/MSMO1 among different species. Rectangles of different colors represent different motifs, and the length of the rectangle represents the number of amino acids.

Fig. 6

Validation of interaction relationships between miR-19b-3p and ELOVL5 and MSMO1. (A) Dual luciferase reporter vectors of ELOVL5 and MSMO1 gene 3′UTRs region. WT indicates the wild type of miR-19b-3p binding sites in ELOVL5 and MSMO1 gene 3′UTRs, MUT indicates the mutant type of miR-19b-3p binding sites in ELOVL5 and MSMO1 gene 3′UTRs. (B) Dual luciferase reporter assay analysis the target relationship of miR-19b-3p with ELOVL5 and MSMO1 in LMH cells. WT indicates wild, MUT indicates mutation, +indicates added, – indicates not added (n = 6). (C) Effect of miR-19b-3p on MSMO1 protein (n = 3).

Fig. 7

Effects of overexpression MSMO1 on TG, TC contents and the expression of related genes. (A) The overexpression efficiency of pcDNA3.1-MSMO1 in LMH. (B) Effects of overexpression MSMO1 on TG and TC contents in LMH. (C) Effects of overexpression MSMO1 on lipid metabolism-related genes. TG indicates triglycerides and TC indicates cholesterol.

Fig. 8

Effects of interfering MSMO1 on TG, TC contents and the expression of related genes. (A) The interfering efficiency of si-MSMO1 in LMH. (B) Effects of interfering MSMO1 on TG and TC contents in LMH. (C) Effects of interfering MSMO1 on lipid metabolism-related genes. TG indicates triglycerides and TC indicates cholesterol.

Fig. 9

miR-19b-3p affects the contents of TG and TC in LMH by targeting MSMO1 and ELOVL5. TG indicates triglycerides and TC indicates cholesterol.

Fig. 10

Effects of estrogen on the expressions of miR-19b-3p and MSMO1. (A) Estrogen downregulates the expression of miR-19b-3p and upregulates the expression of MSMO1 (n = 6). (B) The potential estrogen receptor-binding site located in MSMO1 promoter region. (C) Dual luciferase reporter system verified the estrogen receptor-binding site in MSMO1 (n = 6). (D) Relative expression levels of MSMO1 in LMH treated with different estrogen antagonists (n = 6). WT indicates wild type of the estrogen receptor-binding site, MUT indicates the mutant type of the estrogen receptor-binding site, + indicates added, – indicates not added.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Poult. Sci.
Your Input Welcome
We welcome your input and comments. Please use this form to recommend updates to the information in ZFIN. We appreciate as much detail as possible and references as appropriate. We will review your comments promptly.
Please check the highlighted fields and try again.
Thank you for submitting comments. Your input has been emailed to ZFIN curators who may contact you if additional information is required.
Oops. Something went wrong. Please try again later.