Abstract
Optimization of Cre/LoxP-based assessment of myonuclear accretion. Co-culture of LV-floxed-Luc C2C12 myotubes with Cre C2C12 myoblasts (A–D). A) relative luciferase activity (RLU/protein content) per well 2 days after addition of 0, 2500, 5000 or 10000 myoblasts/cm2 (n = 3). Cells lysed at indicated time points (h) after initiation of co-culturing +/- 10 nM IGF-I (n = 8), B) luciferase activity (RLU) per well, C) protein content (μg/μL) per well, D) relative luciferase activity (RLU/protein content) per well. Co-culture of Cre C2C12 myotubes with LV-floxed-Luc C2C12 myoblasts (E-H). E) relative luciferase activity (RLU/protein content) per well 2 days after addition of 0, 2500, 5000, or 10000 myoblasts/cm2 (n = 3). Cells lysed at indicated time points (hours) after initiation of co-culturing +/- 10 nM IGF-I (n = 8), F) luciferase activity (RLU) per well, G) protein content (μg/μL) per well, H) relative luciferase activity (RLU/protein content) per well. Values are means ± SEM. *p
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