Journal: Molecular Therapy

Article Title: Long Noncoding RNA lncMUMA Reverses Established Skeletal Muscle Atrophy following Mechanical Unloading

doi: 10.1016/j.ymthe.2018.09.014

Figure Lengend Snippet: lncMUMA Expression Associated with Muscle Differentiation in Microgravity-Simulated C2C12 Myoblasts In Vitro and Muscle Mass in HLS Mice (A) Real-time PCR analysis of lncMUMA levels in C2C12 cells with either normal gravity (control) or microgravity-simulated (MGS) culture environment on days 0, 1, 3, 5, and 7 of differentiation. (B) Western blot analysis of MyoD protein level in C2C12 cells with either normal gravity (control) or MGS culture environment on days 0, 1, 3, 5, and 7 of differentiation. (C) Representative images of C2C12 cells with either normal gravity (control) or MGS culture environment on day 7 of differentiation. Myosin was labeled with green fluorescence and the nuclei were labeled with DAPI. Scale bars, 50 μm. (D) The fusion index in C2C12 cells with either normal gravity (control) or MGS culture environment on day 7 of differentiation. (E) Real-time PCR analysis of lncMUMA levels in gastrocnemius muscle of either age-matched control or HLS mice. (F and G) Representative images (F) and muscle mass (G) of gastrocnemius muscle from age-matched control and HLS mice during unloading. Scale bar, 5 mm. n = 5 for in vitro and n = 10 for in vivo . U6 small nuclear RNA is used as the endogenous control of lncRNA. β-actin is used as the endogenous control for protein. Data are presented as mean ± SEM. *p < 0.05 versus the corresponding day 0.

Article Snippet: Following extensive washes with PBS, cells were mounted in mounting medium with DAPI (Merck KGaA) and imaged using the Zeiss Observer Z1 microscope.

Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction, Control, Western Blot, Labeling, Fluorescence, In Vivo

Journal: Molecular Therapy

Article Title: Long Noncoding RNA lncMUMA Reverses Established Skeletal Muscle Atrophy following Mechanical Unloading

doi: 10.1016/j.ymthe.2018.09.014

Figure Lengend Snippet: lncMUMA, but Not Mutated lncMUMA, Counteracted the Decreases in MyoD Protein Level and Myotube Formation in C2C12 Cells Stably Overexpressing miR-762 In Vitro (A) (Left) Western blot analysis of MyoD protein in C2C12 cells stably overexpressing miR-762 transfected with empty vector (EV), lncMUMA vector, and mutated lncMUMA vector (lncMUMA-Mut), respectively, on days 0, 1, 3, 5, and 7 of differentiation. (Right) Representative images of MyoD protein in each group on day 7. (B) Representative images of C2C12 cells in each group on day 7 of differentiation. Myosin was labeled with green fluorescence and the nuclei were labeled with DAPI. Scale bars, 50 μm. (C) The fusion index in C2C12 cells in each group on day 7 of differentiation. n = 5 at each time point for each group. β-actin is used as the control for protein. Data are presented as mean ± SEM. *p < 0.05 versus the corresponding day 0.

Article Snippet: Following extensive washes with PBS, cells were mounted in mounting medium with DAPI (Merck KGaA) and imaged using the Zeiss Observer Z1 microscope.

Techniques: Stable Transfection, In Vitro, Western Blot, Transfection, Plasmid Preparation, Labeling, Fluorescence, Control

Journal: Molecular Therapy

Article Title: Long Noncoding RNA lncMUMA Reverses Established Skeletal Muscle Atrophy following Mechanical Unloading

doi: 10.1016/j.ymthe.2018.09.014

Figure Lengend Snippet: Enforced lncMUMA Expression Could Counteract the Decreases in MyoD Protein Level and Myotube Formation in Microgravity-Simulated C2C12 Myoblasts In Vitro (A) Real-time PCR analysis of lncMUMA levels in C2C12 cells transfected with either lncMUMA overexpression vector (lncMUMA) or empty vector (EV) under microgravity-simulated (MGS) culture environment on days 0, 1, 3, 5, and 7 of differentiation. (B) Western blot analysis of MyoD protein level in C2C12 cells transfected with lncMUMA or EV under MGS culture environment on days 0, 1, 3, 5, and 7 of differentiation. (C) Representative images of C2C12 cells transfected with lncMUMA or EV under MGS culture environment on day 7 of differentiation. Myosin was labeled with green fluorescence and the nuclei were labeled with DAPI. Scale bars, 50 μm. (D) The fusion index in C2C12 cells transfected with lncMUMA or EV under MGS culture environment on day 7 of differentiation. n = 5 at each time point for each group. U6 small nuclear RNA is used as the internal control of lncRNA. β-actin is used as the control for protein. Data are presented as mean ± SEM. *p < 0.05 versus the corresponding day 0.

Article Snippet: Following extensive washes with PBS, cells were mounted in mounting medium with DAPI (Merck KGaA) and imaged using the Zeiss Observer Z1 microscope.

Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction, Transfection, Over Expression, Plasmid Preparation, Western Blot, Labeling, Fluorescence, Control