Figure 4

Astrocyte elevated gene-1 (AEG-1) localizes to nuclear pockets during wound healing in cultured human astrocytes. Confluent astrocyte cultures were scratched to mimic injury and subsequent wound healing by migrating astrocytes was monitored for up to 48 h. Phase-contrast images of the wound tract are shown from 0 to 48 h (A1-F1). In parallel, starting from 8 h post injury, cells were periodically fixed and immunostained with glial fibrillary acidic protein (GFAP) (red, astrocyte marker) and AEG-1 (green) antibodies (A2-E2). Higher resolution micrograph of AEG-1 and GFAP costaining in migrated astrocytes is shown (40 ×, (G)). All images are 20 × original magnification, unless otherwise noted. Cytoplasmic and nuclear protein extracts of astrocytes following wound healing were immunoblotted for AEG-1 (H); cytoplasmic glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and nuclear lamin A/C were used as subcellular fraction normalizing controls (*P <0.01, (J)). Representative data from three individual donors assayed in triplicate. Immunoblotting data is representative of three individual donors.