Figure 1

Ligation of L5-spinal nerve (SNL) induces p38 activation in microglial cells in the spinal cord. (a) Immunostaining of phosphorylated p38 (p-p38) in the spinal cord 3 days after SNL. Dotted line indicates the outline of the spinal cord gray matter. Modified from [34]. (b) p38 kinase assay reveals that SNL increases the phosphorylation of the p38 substrate ATF-2, suggesting an increase in p38 activity. To perform kinase assay, p-p38 was first immunoprecipitated with a p-p38 antibody, then ATF-2 fusion protein was added. Finally, pATF-2 level was detected using Western blotting with a pATF-2 antibody. (c-e) Double immunofluorescence indicates that p-p38 is not colocalized with NeuN, a neuronal marker (c) and GFAP, an astroglial marker (d), but colocalized with OX-42 (CD11b), a microglial marker (e). Modified from [34].