FIGURE 4.

The effect of onset-treatment with SHSST on recruitment/infiltration of residential microglia and peripheral macrophages in the spinal cords from EAE mice. (A–E) Lumbar spinal cord sections were obtained from the sham, EAE, EAE + SHSST, and SHSST groups at day 14–16 post-immunization. The sections were immunostained with anti-Iba-1 antiserum (A–D), and their immunoreactivity was quantified (E). (F) Lysate of the lumbar spinal cord, obtained from each group at day 14–16 post-immunization, was analyzed for mRNA expression of CD11b by real-time PCR. (G,H) Lumbar spinal cords were dissected from each group at day 14–16 post-immunization and analyzed to investigate the degree of recruitment/infiltration of microglia and macrophages by flow cytometry (G); and were quantified (H). CD11b⁺ cells were divided into CD11b⁺/CD45^+(low) cells (R6; microglia) and CD11b⁺/CD45^+(high) cells (R7; macrophage) populations (G), and the percentages of each population are denoted in the graph (H). Bars = 10 μm. Quantified data are expressed as mean inflammation scores, fold induction or % cells ±SEM. (ANOVA testing was performed; ^#p < 0.05 and ^##p < 0.01 versus the sham group; ^∗p < 0.05 and ^∗∗p < 0.01 versus the EAE group.)