Biochemical pathway evaluation

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Efflux pump inhibition (ATPase activity)
The nature of the interaction between the compound and the transporter is determined by ATPase activity assay. Cell lines overexpressing different efflux pumps are incubated with the test compounds and the effect on basal ATPase activity is measured. Compounds that stimulate ATPase are generally considered substrates of the transporter. In the presence of inhibitors, that block the efflux pump, no ATPase activity is detected. ATP consumption is determined by the assay following reported.