Figure 6.

IFNγ is upregulated in spinal cords of ALS mice. (a) The percentage of motoneurons, as identified with VAChT immunostaining on adjacent sections (Supplementary Figure 5a and b), immunoreactive for LIGHT and LT-βR was determined in lumbar spinal cord of wild-type and SOD1^G93A mice at 75, 90 and 110 days (d) of age. (b) Total protein extracts from lumbar spinal cords of wild-type and SOD1^G93A mice at indicated age were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and probed with antibodies to IFNγ and actin (Supplementary Figure 5c). IFNγ signals were quantified, normalized to actin signals and expressed as the ratio of SOD1^G93A to wild-type values. (c) ELISA quantification of IFNγ levels in dissociated spinal cord of 110-day-old wild-type and SOD1^G93A mice and 365-day-old SOD1^WT mice. Values in (a–c) are means±S.D., n=3. (d and f) Immunostainings of wild-type and SOD1^G93A mice lumbar spinal cord sections at 90 days of age using antibody against IFNγ in combination with either GFAP (d), Iba1 (e) or SMI32 (f). Scale bar, 30 μm. (g) Lumbar spinal cord sections of wild-type and SOD1^G93A mice were immunostained as in (f) at 75, 90 and 110 days of age and the percentage of IFNγ immunoreactive motoneurons was determined by counting under fluorescent microscope (75 days, n=3; 90 days, n=4; 110 days, n=5, values are means±S.D.)