Effects of fingolimod on astrocyte activation and glutamate transporter mRNA levels in the spinal cord during EAE.

(A, B) Representative GFAP staining of spinal cord cross sections from a mouse treated with fingolimod in a preventive setting and a sham treated control at the maximum of EAE. Note the reduced GFAP staining after fingolimod treatment. Bar = 100µm (C) Blinded quantification of GFAP immunreactivity as marker of astrocyte activation on spinal cord cross sections. (D) RT-PCR analysis of slc1a2 expression. (E) RT-PCR analysis of slc1a3 expression, n = 6–8 per group. The mRNA expression of a prophylactically treated mouse is set to 1 as reference. All data are given as mean ± SEM. Please note that data are compiled from separate experiments with different starting points of fingolimod application (d0, 11 or 25). The day (d) on the X axis indicates the start of treatment directly after immunization (d0, n = 6 mice per group), at the first sign of symptoms (d11, n = 6 mice per group) or at the early chronic phase of the disease (d25, n = 8 mice per group). Experiments were analysed at the maximum of disease (days 15 or 17 p.i., respectively) for treatment start on day 0 and 11 p.i. and in the later chronic phase of EAE (day 80 p.i.) after treatment start on day 25 p.i. * p < 0.05, ** p < 0.01; *** p < 0.001, Mann Whitney test.