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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Mol Microbiol. 2014 Jan 21;91(5):965–975. doi: 10.1111/mmi.12508

Figure 5.

Figure 5

Western blot analyses to determine the in vivo stability of a labile TolC protein carrying P246R and S350C alterations in its mature sequence. Protein extracts from approximately 5 × 107 cells grown overnight at 37°C were analyzed by SDS-PAGE and electro-transferred onto PVDF membranes. Membrane blots were probed with HisProbe-HRP to detect His-tagged TolCP246R, S350C. HisProbe-HRP also recognized an unknown band, which was used as a loading control (LC). Protein levels were quantified with Quantity One software (Bio-Rad). TolCP246R, S350C levels were determined relative to LC and then normalized to the wild type (AcrAB+) value of 1.