After completion of muscle strength measurements experiments were terminated by excision of the injured left soleus muscle for subsequent histology and immunohistochemistry.
Myofiber necrosis is known to induce a rapid and transient activation of the innate immune system [ 29], which is essential for successful completion of muscle regeneration.
Thus, the down-regulation of genes involved in this pathway within the FFDF cluster suggests the completion of muscle developmental processes dependent on satellite cell formation.
Furthermore, single-nucleus RFP+ cells could be detected at the niche of endogenous MuSCs after completion of muscle regeneration, suggesting that the transplanted MuSCs are capable of proper homing.
Single-nucleus RFP+ cells could also be detected at the niche of endogenous MuSCs after completion of muscle regeneration, suggesting that the MuSCs expanded in cytokine medium are capable of proper homing after transplantation.
After completion of the muscle strength measurements, the animals were killed with an overdose of anesthetic and the soleus muscles were fixed in buffered paraformaldehyde (4%) for histological examination.
Functional Groups 3 and 4 contained terms linked to neurogenesis, which may correspond with completion of the muscle innervation process that began with secondary myotube formation [ 11].
Fibrotic lesions were highly proliferative in 2-week-old mutants, whereas the fibrotic lesions of older mutants showed little proliferation indicating the completion of local muscle replacement by scar tissue.
At the completion of the experiment, muscle tissue was dissected from the right posterior appendage and the abdomen (similar to above samples) and frozen in separate scintillation vials in order to determine if SIR values vary for these different tissue types.
EMG data was normalised to maximum voluntary isometric contractions (MVIC); three MVICs were recorded for each muscle following completion of walking trials.
