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. 2000 Mar 14;97(6):2515–2520. doi: 10.1073/pnas.97.6.2515

Figure 5.

Figure 5

Effect of cyclosporin A and cis(Z)-flupentixol on the IAAP labeling of Pgp in the Vi-trapped and native conformations. (A) The effect of cyclosporin A and cis(Z)-flupentixol on the IAAP labeling of control and Pgp treated with 1.2 mM 8AzidoATP and 250 μM Vi at 37°C for 10 min immediately after trapping. After incubation at 37°C, the samples were centrifuged at 300,000 × g for 10 min at 4°C and resuspended in ice-cold ATPase assay buffer. The samples then were immediately incubated with cyclosporin A (1 μM) or cis(Z)-flupentixol (25 μM) for 3 min at room temperature, with IAAP (5 nM) for an additional 3 min, and then photocrosslinked as described in Materials and Methods. (B) Same experimental conditions as A except that the samples were incubated with 1.2 mM ATP for 15 min at 37°C before treatment with cyclosporin A or cis(Z)-flupentixol and labeling with IAAP. The various treatments are given above each autoradiogram.