Fig. 6

Localization of C9ORF72 over development in vitro. Immunofluorescence was carried out on primary cultured cortical neurons. a At 1 DIV, C9ORF72 (red) labeling was present within cell bodies, excluding nuclei (DAPI, blue) and punctate localization was present in neurites and growth cones (β-III tubulin, green). b Co-staining with phalloidin (green) at 3 DIV confirmed localization of C9ORF72 (red) labeling to growth cones and to filopodia (arrows). c At 14 DIV, C9ORF72 (red) was localized to nuclei of a population of neurons (arrows) but was less intensely expressed in nuclei of other neurons (arrowhead). Neurons indicated by MAP2 (green). Neurons with nuclear immunolabeling for C9ORF72 frequently had punctate somal localization of this protein. Inset (c) shows C9ORF72 labeling in nuclei and in puncta in surrounding cytoplasm. Scale bar: A, 2.5 μm; B, C, 10 μm