Fig. 4. Cellular localization of SHP1 mRNA and protein 3 d after transection of the facial nerve and 1 d after direct cortical lesion. A, Light micrographs of hematoxylin-stained in situ hybridization sections show that silver grains accumulate over small, darkly staining nuclei (red arrows) in the axotomized facial nucleus, whereas motoneurons remain unlabeled. B, C, Facial motor nuclei 3 d after facial nerve transection, triple labeling for Cy3 (anti-SHP1, red), Cy5 (B, microglia, anti-Ox42; green), and DTAF (C, astrocytes, anti-GFAP; green). After transection of the facial nerve, SHP1-IR (red) colocalizes (red and green overlap resulting in yellow) with Ox42-IR on activated microglia (B, arrowheads) and GFAP-IR on astrocytes (C, arrows). D, After direct cortical lesion, invading macrophages identified by anti-ED-1-antibody (green) and a subpopulation of T-lymphocytes identified by anti-CD3-antibody (green) are also positive for SHP1-IR (red, colocalization in yellow).N, Neuron; Con, control, unoperated side;Ax, axotomized side; Dcl, direct cortical lesion side. Scale bars, 100 μm.