Sp2509

Rabbit monoclonal antibodies raised against LSD1, ATG7, di-methyl-histone H3 (Lys4), histone H3, ubiquitin, and PARP were purchased from Cell Signaling Technology (Danvers, MA, USA); the anti-GAPDH antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-Flag and anti-HA monoclonal antibodies were from Sigma (St. Louis, MO, USA); the anti-p62 rabbit polyclonal antibody was from GeneTex (San Antonio, TX, USA); the anti-LC3 rabbit polyclonal antibody was from Novus Biologicals (Littleton, CO, USA); the anti-methylated lysine rabbit polyclonal antibody was from Abcam (Cambridge, MA, USA). The LSD1 inhibitor SP2509 was obtained by Medchemexpress (Monmouth Junction, NJ, USA). All chemicals including chloroquine were purchased from Sigma, unless otherwise indicated. GFP-LC3 was kindly provided by Dr. Jennifer Leppincott-Schwartz (National Institutes of Child Health and Human Development, Bethesda, MD, USA).

Working dilutions consisted 20% cremaphor (Sigma, St. Louis, USA), 20% dimethyl sulfoxide (Sigma, St. Louis, USA) and 60% phosphate-buffered saline (PBS). SP2509 was obtained from Med Chem Express (New Jersey, USA). Stock solutions were prepared by dissolving the components in the working dilutions with a concentration of 5 mg/ml.
Rats were separated randomly in three groups (n = 10 for each group), including sham-operated, SCI-only, and Add-SP2509 groups: one was treated with 25 mg/kg SP2509 immediately and 48 h after the wound was closed, while the other two groups were treated with 5 ml/kg working dilutions^{20 (link),44 (link)}.