Supplementary Figure 2: Immunofluorescence for TNFα in EAE remission.
From: Maladaptive cortical hyperactivity upon recovery from experimental autoimmune encephalomyelitis
Coronal slices from remission animals were stained with the following antibodies to label specific cell types for idenfication of the source of TNFα: (a) astrocytic anti-EAAT1, (b) microglial anti-Iba-1, (c) neuronal marker anti-NeuN, (d) anti-GAD67 for GABAergic neurons and (e) anti-CamKII for CamKII+ excitatory neurons; DAPI was used for nuclear staining. (Center) TNFα was stained with anti-TNFα (green), (Right) Merged images of respective stainings are shown. (Scale bars = 30 µm; arrow heads indicate cells which do not express TNFα, arrows indicate cells which are positive for TNFα.) (f) (Left) Magnification of staining for TNFα, scale bar = 1 µm. (Right) Superresolution image (resolution 36 nm) showing the intracellular compartment, which was delineated by CamKII staining (red). Remarkably, TNF alpha (green) immunosignal was present in the intracellular compartment as well as in the extracellular compartment, which is in line with neuronal TNFα secretion. Representative images from 5 animals, scale bar = 500 nm.
