Fig. 5

Caspase-inhibited neurons display PS-translocation. Neurons were incubated with colchicine (1 µM) and N-benzyloxycarbonyl-Val-Ala-aspartyl-fluoromethyl ketone (zVAD-fmk; 100 µM) for 24 hr and stained with fluorescent annexin V [phosphatidylserine (PS)-translocation], calcein acetoxymethyl ester (calcein-AM; plasma membrane integrity) and H-33342 (chromatin structure). Fluorescence images were collected by confocal microscopy (63×, NA 1.32 lens). Top: xy-section through the cell bodies. Arrows indicate condensed nuclei and an asterisk indicates an annexin V-positive neuron. The width of the scale bar corresponds to 10 µm. Middle: xy-section through the annexin V-positive neuron in higher magnification. Bottom: transversal section (xz-plane) demonstrates that the entire surface of the neuron was surface stained with annexin V. The width of the scale bar corresponds to 9 µm. Data are representative of five experiments in three independent cell preparations.