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. 2020 May 13;27(12):960–975. doi: 10.1038/s41417-020-0178-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a T7EI assay of mice genes produced by intramuscular injections of rAAV–SaCas9. Numbers of mutant muscle tissues with bands of 168, 338 and 506 bp in AAV-sgMstn2, RAAV-sgMstn2 and only one band of 506 bp in AAV-CN. The red arrow indicates the band of enzyme digestion. b Stacked histogram showing the percentage distribution of indels at Mstn in muscle tissues of mice, as measured by sequencing analyses. Data are presented as mean ± SEM from five mice in each group. c The muscle tissues in AAV-sgMstn2, RAAV-sgMstn2 and AAV-CN groups analyzed by western blot using an anti-MSTN antibody. And anti-GAPDH was used as loading control. d Stacked histogram showing the percentage distribution of MSTN expression in different muscle tissues. Data represent means ± SEM (The number of samples was 3 per group, and each sample was repeated three times). *: 0.01 < P < 0.05, **P < 0.01 when AAV-sgMstn2, RAAV-sgMstn2-treated were compared with AAV-CN-treated. e Changes of left thigh muscle mass in the aged mice in AAV-CN, AAV-sgMstn2 and RAAV-sgMstn2 groups. Compared with the AAV-CN-treated group, the muscle mass of the MSTN gene knockout group was significantly increased by AAV-sgMstn2 and RAAV-sgMstn2-treated groups (in red cycle). f Changes in body weight from one to eight weeks in different experimental groups. Data represent means ± SEM, N = 10 per group. g, h The weight of the quadriceps and adductor muscle in the different groups at 8 week. Data represent means ± SEM, N = 10 per group. *: 0.01 < P < 0.05, **P < 0.01, when AAV-sgMstn2, RAAV-sgMstn2-treated were compared with AAV-CN-treated.