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. 2020 Apr 23;11:556. doi: 10.3389/fmicb.2020.00556

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Copyright © 2020 Feng, Liu, Wang, Wang, Zang, Liu, Song, Gu, Fan, Yang, Chen and Cui.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Inhibition of tetracycline (TC) transport activity by ortho-vanadate, CCCP, and MDR substrates. (A) ortho-vanadate (1 mM) inhibited efflux of tetracycline (TC). (B) CCCP (40 μM) inhibited efflux of tetracycline (TC). CCCP had a stronger inhibitory effect on tetracycline efflux than ortho-vanadate. (C) Kinetic analysis of the inhibitory effect of ortho-vanadate on tetracycline (TC) efflux activity. The assay was carried out in the presence of increasing concentrations of ortho-vanadate ranging from 0 to 2.5 mM. (D) Kinetic analysis of the inhibitory effect of CCCP on tetracycline (TC) efflux activity. The experimental conditions were the same as for panel C. The assay was carried out in the presence of increasing concentrations of CCCP ranging from 0 to 160 μM. (E) Kinetic analysis of the inhibitory effect of reserpine on tetracycline (TC) efflux activity. The assay was carried out in the presence of increasing concentrations of reserpine ranging from 0 to 80 μM. (F) Kinetic analysis of the inhibitory effect of EB on tetracycline (TC) efflux activity. The assay was carried out in the presence of increasing concentrations of EB ranging from 0 to 20 mM. (G) Kinetic analysis of the inhibitory effect of Hoechst33342 on tetracycline (TC) efflux activity. The assay was carried out in the presence of increasing concentrations of Hoechst33342 ranging from 0 to 1.0 μM. (H) A table showing a summary of the IC 50 values. The IC 50 values were calculated as described under Materials and methods.