Molecular Therapy - Nucleic Acids

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Molecular Therapy - Nucleic Acids Reverse Expression of Aging-Associated Molecules through Transfection of miRNAs to Aged Mice Jung-Hee Kim, Bo-Ram Lee, Eun-Sook Choi, Kyeong-Min Lee, Seong-Kyoon Choi, Jung Hoon Cho, Won Bae Jeon, Eunjoo Kim Molecular Therapy - Nucleic Acids Volume 6, Pages 106-115 (March 2017) DOI: 10.1016/j.omtn.2016.11.005 Copyright © 2016 The Author(s) Terms and Conditions

Figure 1 The Top Network Associated with the DERs Selected by the Criteria of 4-fold Change in Whole Blood The functions related to this network are defined as cancer, organismal injury and abnormalities, and reproductive system disease. miRNAs, indicated as green half circles, represent the selected DERs. The connection with Ppara is shown using pink arrows. The analysis was performed using the Ingenuity Pathway Analysis (IPA) program at http://www.ingenuity.com. Error bars show SD. Molecular Therapy - Nucleic Acids 2017 6, 106-115DOI: (10.1016/j.omtn.2016.11.005) Copyright © 2016 The Author(s) Terms and Conditions

Figure 2 Functional Changes Caused by DERs Identified in the Whole Blood of Aged Mice (A) Disease states are expressed as probability of significance, calculated as p < 0.05 by IPA analysis. (B) Histological analysis of fatty liver according to age in mice. The deposited fats were observed in empty vacuoles (red arrows); thus, an elevated number of vacuoles implies the development of fatty liver. Error bars show SD. Molecular Therapy - Nucleic Acids 2017 6, 106-115DOI: (10.1016/j.omtn.2016.11.005) Copyright © 2016 The Author(s) Terms and Conditions

Figure 3 Expression Level of miRNAs and Proteins in Whole Blood and Liver According to Aging Stage (A and B) Expression is shown of differentially expressed miRNAs in (A) whole blood and (B) liver tissue of mice, assessed by qPCR and according to age. (C) Expression of proteins involved in the miRNA network was analyzed by western blotting. Red asterisk(s), comparison to 8-month-old mice; black asterisk(s), comparison to 3-month-old mice (*p < 0.05, **p < 0.01, and ***p < 0.001). Error bars show SD. Molecular Therapy - Nucleic Acids 2017 6, 106-115DOI: (10.1016/j.omtn.2016.11.005) Copyright © 2016 The Author(s) Terms and Conditions

Figure 4 Alteration of miRNA and Protein Level Induced by Six miRNAs in Liver (A and B) Changes in (A) miRNA and (B) protein levels in the liver of 18-month-old mice after intraperitoneal injection of six miRNAs (*p < 0.05, **p < 0.01, and ***p < 0.001). (+) miRNA indicates the expression level in mice in the group injected with the six miRNAs. Error bars show SD. Molecular Therapy - Nucleic Acids 2017 6, 106-115DOI: (10.1016/j.omtn.2016.11.005) Copyright © 2016 The Author(s) Terms and Conditions

Figure 5 Expression of Aging-Associated Molecules in Livers Transfected with Six miRNAs (A and B) The (A) mRNA levels of telomerase-related genes and (B) expression of proteins involved in aging pathways. (+) miRNA indicates the expression level in mice in the group injected with the six miRNAs. Error bars show SD. Molecular Therapy - Nucleic Acids 2017 6, 106-115DOI: (10.1016/j.omtn.2016.11.005) Copyright © 2016 The Author(s) Terms and Conditions