For western blotting and quantitative PCR (qPCR), fresh spinal cord and muscles were removed and snap frozen.
Interventions: Rat anterior tibial and soleus muscles were removed and repetitively stretched about 20% of the muscle length by using the Magnus method in Krebs-Ringer solution.
Breast muscles were removed and cut into 3 parts and stored at 0 °C for 0 h, 3 h and 10 h.
The frozen muscles were removed with an electric dental drill in liquid nitrogen, and processed for freeze-substitution fixation in acetone containing 0.2% glutaraldehyde at −80 °C for 24 h and then −30 °C and 4 °C for 2 h each.
All affected components including subcutaneous tissue, fascia, and muscles were removed and sent for histopathologic diagnosis of necrotizing fasciitis to include in the study.
Tibialis anterior, gastrocnemius, quadriceps, soleus, extensor digitorum longus and diaphragm muscles were removed to perform molecular (enzymology, RT-qPCR and western blotting) and histological analyses.
The muscles were removed from the lumbar spines, while the posterior bony elements, capsular structures and ligaments were left intact (Baranto et al. 2005a; Baranto et al. 2005b).
Excess muscles were removed, and joints were fixed in a 10% neutral buffered formalin solution (Thermo Fisher Scientific, MA, USA) for 14 days at room temperature.
The brain and major muscles were removed and the cavities filled with clay, vegetable matter and ash.
Soleus muscles were removed from anesthetized animals, frozen in liquid nitrogen, and stored at − 80°C.
Briefly, adult mice were fixed with paraformaldehyde; the muscles were removed, post-fixed, rinsed and mounted on slides.
