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. 2020 May 15;48(13):7005–7017. doi: 10.1093/nar/gkaa346

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Deletion and overexpression of YcaQ sensitizes E. coli to mechlorethamine. (A) Colony dilution assay for E. coli deletion strains exposed to increasing concentrations of mechlorethamine (HN2). Values are mean ± SEM (n = 3). Percent (%) survival is relative to untreated cells. (B) EC₅₀ values derived from data in panel A. Significance values were calculated using a one-way ANOVA (*P = 0.0140; **P = 0.0085; ***P < 0.0001; n.s, not significant). (C) Colony dilution assay for E. coli strains complimented with YcaQ or empty vector. Values are mean ± SEM (n = 3). (D) Quantification of the data shown in panel D. One-way ANOVA values: ***P = 0.0004, n.s., not significant. (E) Colony dilution assay showing the effect of mechlorethamine (HN2) on wild-type E. coli overexpressing YcaQ variants and/or EndoIV. (F) Quantification of the data shown in panel E. One-way ANOVA significance values: *P = 0.0088; **P = 0.0013; ***P < 0.0001; n.s, not significant. (G) MMS EC₅₀ values (mM) for various E. coli deletion (Δ) or over-expression (OE) strains. (H) qRT-PCR results of DNA repair genes after treatment of E. coli with 5 mM MMS or 200 μM mechlorethamine (HN2) for 2 hr. Average ± SEM for three biological replicates.