Figure 6
Flow cytometric analysis of immune cells from the spinal cord and SNs separated with Percoll centrifugation. (a) Total cell lysates of spinal cord immune cells enriched with Percoll centrifugation were subjected to immunoblotting with the anti-KS antibody. (b) Immune cells purified from SNs and spinal cords were subjected to flow cytometry analysis. Representative results are shown (n=3). Gate A corresponds to lymphocytes, gate B corresponds to microglia/macrophages (c). The number of lymphocytes and macrophages were counted in normal and EAN SNs. (d) The number of lymphocytes and microglia/macrophages were counted in normal and EAN spinal cords. (e) Expression of CD45 was analyzed by flow cytometry. Red line and blue line correspond to control and EAN peak, respectively. (f) Mean fluorescent intensity of CD45 was calculated in microglia/macrophages (gate B). (g) KS expression in the microglia/macrophages (gate B) in the spinal cords was measured. (h) Mean fluorescent intensity of KS was calculated in lymphocytes (gate A) and microglia/macrophages (gate B) in the spinal cords. (i) KS expression in the gated lymphocytes and microglia/macrophages were calculated in normal and EAN spinal cords. (j) KS expression in the gated lymphocytes and macrophages were calculated in normal and EAN SNs. Open columns, normal rats; and closed columns, EAN rats. Each column represents average±S.D. The P-value was calculated using Student's two-tailed t-test. *P<0.05, **P<0.01. N.S., not significant
