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. 1997 Dec 1;139(5):1109–1118. doi: 10.1083/jcb.139.5.1109

Figure 5.

Figure 5

NAD+ is not required for BFA-induced coatomer dissociation from the Golgi complex in permeabilized cells. Intact RBL cells (a and b) were treated with 3 μg/ml BFA (b), or were permeabilized with 3 U/ml SLO and then exposed to control buffer (c), or to a buffer containing 150 μM NAD+ (e), or 10 μg/ml BFA alone (d), or BFA in combination with 150 μM NAD+ (f). The cells were fixed, permeabilized with saponin, and stained with anti–β-COP antibody. SLO permeabilization induces a partial detachment of β-COP from Golgi complex (c) compared to intact cells (a), but BFA is completely effective in inducing the total cytosolic redistribution of β-COP independently of the presence of NAD+ in the permeabilization buffer (d and f). Pre–ADP-ribosylated cytosol (g and h) behaves indistinguishably from control cytosol. Similar results were obtained in permeabilized CHO cells (not shown). The experiments were repeated four times in duplicate with similar results. Bar, 5 μm.