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  Indian J Med Microbiol
 

Figure 3: Effect of curcumin on basal and verapamil-stimulated ATPase activity and photoaffinity labeling of MDR1 with 8-azido-ATP-biotin: (a) Y79-MDR1 cells expressing P-gp (100 μg of protein/ml) were incubated with increasing concentration of curcumin in the presence of verapamil (◼) or an equivalent volume of DMSO (♦) in the ATPase assay buffer. Values are mean ± SE from at least 3 independent experiments. (b) Crude membranes (100 μg protein) were incubated with 100 μM 8-azido-ATP-biotin in the dark on ice for 5 min in the presence and absence of 10 μM curcumin. The protein was electrophoresed, blotted and visualized with streptavidin horseradish peroxidase. The addition of 10 mM ATP prevented photo-labeling. Lane 1: 8-azido-ATP-biotin, Lane 2: 10 μM curcumin and Lane 3: 10 mM ATP

Figure 3: Effect of curcumin on basal and verapamil-stimulated ATPase activity and photoaffinity labeling of MDR1 with 8-azido-ATP-biotin: (a) Y79-MDR1 cells expressing P-gp (100 μg of protein/ml) were incubated with increasing concentration of curcumin in the presence of verapamil (◼) or an equivalent volume of DMSO (♦) in the ATPase assay buffer. Values are mean ± SE from at least 3 independent experiments. (b) Crude membranes (100 μg protein) were incubated with 100 μM 8-azido-ATP-biotin in the dark on ice for 5 min in the presence and absence of 10 μM curcumin. The protein was electrophoresed, blotted and visualized with streptavidin horseradish peroxidase. The addition of 10 mM ATP prevented photo-labeling. Lane 1: 8-azido-ATP-biotin, Lane 2: 10 μM curcumin and Lane 3: 10 mM ATP