Fig. 7
Effects of 1-oleoyl-LPA, an agonist of LPAR1/2, on neurological symptoms and main pathophysiological features in spinal cords of mice after EAEhigh induction. A, B Following immunization, neurological symptoms in sham, EAEhigh, and EAEhigh + 1-oleoyl-LPA (0.5 and 1 mg/kg) groups were measured daily (A) and scores from days 11–18 were summed (B). C At 19 days after immunization, cryosections (n = 3 per spinal cord) of lumbar spinal cords (n = 3 per group) were stained with LFB (upper panel), H&E stain (middle panel), or immunochemical stained with Iba-1 antibody (lower panel). D–H, K–M At day 19, lysates of lumbar spinal cords (n = 3) from each group were analyzed by Western blot to measure protein expression levels of MBP (D), Iba-1 (E), COX-2 (F), PECAM-1 (G), 4-HNE (H), LPAR1 (K), LPAR2 (L), and LPAR3 (M). I, J Total RNA (n = 3 per group) was extracted and used for real-time PCR to measure mRNA expression levels of NOX2 (I) and NOX-3 (J). Data are expressed as mean expressive value ± SEM (ANOVA test; #p < 0.05 and ##p < 0.01 versus sham group; *p < 0.05 and **p < 0.01 versus EAEhigh group)