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Efflux pump inhibition assays were performed following previously published protocols and modified using M. aurum cells9,15,31. The effect of the synthesized compounds and verapamil (positive control) on the accumulation of ethidium bromide (EtBr) was determined by measuring fluorescence using a fluorimeter (FLUOstar OPTIMA, BMG Labtech) and fluorescence data was acquired every 60 s for a total period of 60 min. The compounds were used at one quarter of their MICs and EtBr (a known efflux pump substrate) at a concentration of 1.3 µM (0.5 mg L−1).
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