Disturbance of mTOR signaling in vivo in GAA‐KO mice

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Disturbance of mTOR signaling in vivo in GAA‐KO mice Disturbance of mTOR signaling in vivo in GAA‐KO mice Muscle biopsies (white part of gastrocnemius) were obtained from 4‐ to 6‐month‐old WT and GAA‐KO (KO) mice. A, BWestern blot analysis of whole muscle lysates from WT and GAA‐KO mice with the indicated antibodies. Graphical presentation of the data is shown in (B). Data illustrate the mean ± SE. n = 6 for p‐4E‐BP1/4E‐BP1, p‐S6/S6, p‐PRAS40/PRAS40, and p‐TSC2/TSC2; n = 5 for p‐AMPKα; n = 3 for LKB1. *P < 0.05, **P < 0.01, ***P < 0.001, Student's t‐test. GAPDH was used as a loading control.CAn increase in ADP/ATP ratio in whole muscle of GAA‐KO mice. Data illustrate the mean ± SE. n = 3 for WT; n = 4 for KO. *P < 0.05, Student's t‐test.D, EMuscle tissues derived from WT (n = 3) and GAA‐KO mice (n = 4) were homogenized in lysis buffer and subjected to fractionation to obtain lysosome‐enriched fractions. The isolated fractions were then examined by Western blot showing increased levels of total mTOR, TSC2, and AMPKα in GAA‐KO. Graphical presentation of the data is shown in (E). Graphs represent mean ± SE. **P < 0.01, ***P < 0.001, Student's t‐test. RHEB and Ponceau S staining were used to verify equal protein loading.FImmunostaining of a single fiber from a GAA‐KO mouse with anti‐LAMP1 (red) and anti‐mTOR (green) antibodies showing extensive co‐localization of the two stains. Scale bar: 10 μm.Source data are available online for this figure. Jeong‐A Lim et al. EMBO Mol Med. 2017;9:353-370 © as stated in the article, figure or figure legend