Figure 3: Molecular characterization of viable muscle stem cells and redox status.

(a, b) Real-time quantitative PCR of satellite cells (n=5 animals). (c–h) NF-κB activity in post mortem and anoxic satellite cells (in all experiments n=5 mice). (c, d) Histology of normal muscle showing lacZ expression only in endothelial cells and some fibroblastic interstitial cells (blue nuclei) (haematoxylin and eosin (H&E) staining) using both p105lacZ transgenic mice (c) and Igkappa-lacZ (d). (e, f) 4 days post mortem, (g, h) 8 days post mortem, note reduction in lacZ-expressing cells, and no expression in cells in satellite cell position adjacent to myofibres. (i) Comparison of oxygen consumption between cells extracted 0 or 4 days post mortem after 30 min or 12 h in culture (n=5 mice). (j) Level of ATP in day 0, 4 and 8 post mortem satellite cells determined by fluorescence using luciferase activity (n=5 mice). (k) Determination of the ability to transform energy and the redox state ratio between NAD and NADH in cells freshly isolated or extracted 4 days post mortem (n=5 mice). (l) Measurement of the level of ROS in satellite cells expressed as a ratio between day 0 and 4 days post mortem (n=5 mice). (m) Enumeration of satellite cells at 0 and 4 days post mortem after variation of antioxidants in the satellite cell environment, by treating Tg:Pax7-nGFP mice with N-acetyl-L-cysteine (NAC) or buthionine sulfoximine (BSO) (n=5 mice). Statistical analysis was performed using paired or unpaired Student's t-tests, a minimum of 95% confidence interval for significance; P-values indicated on figures are <0.05 (*), <0.01 (**), and <0.001 (***). Figures display average values of all tests ±s.e.m.