Sham-2
A total of 3mm tissue from both ends of the SCI site was collected, immediately frozen in liquid nitrogen, and stored at -80. Spinal cord tissue was lysed with 300 l of TRIzol (#15596026, Thermo-Fisher Scientific) with pipetting. After mixing with chloroform and centrifugation, the RNA-containing aqueous supernatant was transferred to a new tube. To facilitate RNA precipitation, 0.5 l of 20 g/l glycogen (#10814010, Thermo-Fisher Scientific), 40 l of 2 M sodium acetate, and 500 l of 100% ethanol were sequentially added. This crude RNA extract was stored at 80 C. For further purification, the sample underwent a series of clean-up steps, DNaseI treatment, and RNA elution according to the Ambion RecoverAll Nucleic Acid Isolation Kit (#AM1975, Thermo-Fisher Scientific) instructions. All RNA samples were stored in LoBind microcentrifuge tubes (#22431021, Eppendorf AG).RNA purity was checked using the kaiaoK5500 Spectrophotometer (Kaiao). RNA integrity and concentration were assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies).