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Western blot analysis to determine AcrA levels. Protein extracts from approximately 5 × 107 cells grown overnight at 37°C were analyzed by SDS-PAGE and electro-transferred onto PVDF membranes. Membranes were blotted with primary antibodies against AcrA and LamB. LamB was used as a gel loading control. Protein levels were quantified with Quantity One software (Bio-Rad). AcrA levels were determined relative to LamB and then normalized to the wild type value of 1.
