Eccentric contraction increases hydrogen peroxide levels and alters gene expression through Nox2 in skeletal muscle of male mice

Control, concentric contraction, eccentric contraction, and eccentric + Nox2 inhibitor groups were established with 3 mice each. The tibialis anterior muscle of the left leg of the mice was electrically stimulated using a nerve electrode to the peroneal nerve under triadic anesthesia. Electrical stimulation was performed 50 times at 100 Hz, once every 3 seconds for 700 ms. ECC was performed 200 ms after electrical stimulation, followed by stretching using exercise equipment. 20 min before ECC, the Nox2 inhibitor GSK2795039 was administered to the tibialis anterior muscle. nucleospin RNA (Takara Bio) was used to analyze the mouse anterior Total RNA was extracted from the tibialis muscle. mRNA was purified from total RNA using Poly-A selection and mRNA samples were checked for quality using a Byoanalayzer (Agilent). 150 base pair paired-end sequencing reads were run using standard Illumina protocols on NovaSeq6000 (Illumina).