Additional file 3: of Activation of GPR40 produces mechanical antiallodynia via the spinal glial interleukin-10/β-endorphin pathway

Figure S3. Stimulatory effect of intrathecal injection of GW9508 on β-endorphin expression in the spinal dorsal horn of neuropathic rats induced by L5/L6 spinal nerve ligation. The spinal lumbar enlargements were obtained 1 hour after intrathecal injection of saline (10 μl) or GW9508 (30 μg). For the gene and protein analysis study, expression of the β-endorphin precursor POMC gene (A) and β-endorphin protein (B) levels were determined using qRT-PCR and a specific fluorescent immunoassay kit, respectively. For the immunostaining study, the spinal lumbar enlargements were frozen. Immunofluorescence was stained with the β-endorphin antibody and photomicrographs were taken from the entire spinal cord section (C, D, 500 μm). E. The immunolabeled surface areas of β-endorphin from the spinal dorsal horn laminae I-V indicated in white lines were quantified using the ImageJ program. Data are presented as mean ± SEM (N = 5~8 per group). * P < 0.05, vs saline group; analyzed by unpaired and two-tailed Student t-test. (ZIP 464 kb)