Figure 3

Western blot and immunohistochemical detection of anti-neuroinflammatory events in the brainstem and spinal cord. The western blot image is representative of three independent experiments. A representative blot of Iba-1 (A) or TNF-α (B) demeonstrates a significant reduction in the expression of these molecules in the brainstem and spinal cord of melittin-treated hSOD1^G93A mice. The expression of MAP2 (a neuron marker) is dramatically increased in both the brainstem and spinal cord of melittin-treated hSOD1^G93A mice compared with saline-treated hSOD1^G93A mice (C). α-tubulin was used for loading control. The activation of p38 is increased in the brainstem and spinal cord of hSOD1^G93A mice, but melittin-treated hSOD1^G93A mice showed reduced expression of phospho-p38 (E). The expression of p38 was used as a comparison for the expression of activated phospho-p38. Selected free-floating fifth spinal cord or brainstem section (40 μm) from saline- (n= 3) or melittin-treated hSOD1^G93A mice (n= 4) was stained with an anti-Iba-1. The immunohistochemical study of Iba-1 in the facial nucleus of the brainstem and the anterior horns of the lumbar (L4) spinal cord confirmed that melittin treatment reduced the number of Iba-1-expressing cells in the hSOD1^G93A mice compared to age-matched control mice (G-J). The large box presents higher magnification views of the small box in the facial nucleus region of the brainstem in control hSOD1^G93A mice (G). Scale bars = 200 μm (G, H). Scale bars = 100 μm (I, J). MT: melittin, BS: brainstem, SP: spinal cord.