A11319

After pain behavioral tests, the rats were anesthetized and sacrificed by decapitation on day 12 after inoculation of MRMT-1 cells. Lumbar spinal cord was dissected and then homogenized in ice-cold radioimmunoprecipitation assay (RIPA) lysis buffer containing a cocktail of protease inhibitors (Sigma). After centrifugation at 12,000 g for 15 min, supernatant was used for Western blot analysis. Protein concentrations were determined by bicinchoninic acid assay method. Equal amounts of protein samples were separated in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were transferred onto a polyvinylidene difluoride membrane and then incubated with the appropriate primary antibodies at 4°C overnight. The following antibodies were used in this study and ABclonal Technology (Wuhan, China): mouse rabbit anti-Drp1 (1:1000, A2586), anti-OPA1 (1:1000, A9833), mouse anti-glial fibrillary acidic protein (GFAP) (1:1000, A0014), rabbit anti-caspase 3 (1:1000, A11319), rabbit anti-Bcl-2 (1:1000, A0208), rabbit anti-β-actin (1:5000, AC004). Horse radish peroxidase (HRP)-conjugated secondary antibodies (1:5000, Abcam) were used to visualize the primary antibodies. Infrared Imaging System (Gene Company Limited, Hongkong, China) was applied to detect immunoreactive bands.