Figure 6From: Histone H4 deacetylation plays a critical role in early gene silencing during neuronal apoptosisHDAC3 translocates to the nuclei of apoptotic cells. Retinal sections from eyes before optic nerve crush (ONC) and at several points after ONC were double labeled with antibodies against HDAC3 (green) and γH2AX (red), to identify apoptotic cells (see Figure 5). (A) Cells in the ganglion cell layer prior to ONC exhibited diffuse cytoplasmic HDAC3 labeling and a densely labeled spot of γH2AX (stage I labeling), which associated with the nucleoli (arrows). Representative unlabeled nuclei are indicated (N). (B) Following ONC, HDAC3 labeling began to appear in the nuclei (asterisk in merged image), while γH2AX labeling was detectable as a cytoplasmic perinuclear ring (stage II labeling, arrowhead). (C) HDAC3 labeling continued to increase in the nuclei (asterisks), while the overall intensity of γH2AX labeling increased in both the cytoplasm (arrowhead) and the nuclei, which appear yellow in the merged image (asterisks). (D) At later time points post ONC, a greater proportion of cells exhibited nuclear colocalization of HDAC3 and γH2AX (stage III labeling, arrowhead). (E, F) HDAC3 and γH2AX labeling continued to colocalize (asterisks), although nuclei of dying cells were less well defined. Scale bar = 5 μm.Back to article page