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. 2022 Dec 8;50(21):12355–12368. doi: 10.1093/nar/gkac1124

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© The Author(s) 2022. 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 (https://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 1. — Effects of RM plasmid copy number on intracellular concentrations of Esp1396I enzymes and levels of protection from phage infection. (A, B) Images of cells with an empty pBR322 plasmid vector (‘pMid’), and cells transformed with indicated Esp1396I_fluo plasmids are shown in transmitted light (TL), Venus (yellow), and mCherry (magenta) channels. Scale bar is 20 μm. The contrasts in (A) and (B) are different to reveal the differences in fluorescence signals. (C–E) The distributions of REase (C) and MTase (D) molecule numbers and of MTase to REase ratios (E) in individual cells carrying the Esp1396I_fluo system on indicated plasmids. (F) Protection levels of cells carrying indicated RM plasmids or the pMid vector from phage λ_vir infection are shown. Protection levels were calculated by dividing the titer of phage lysate on the lawn of cells carrying the pMid vector by a titer determined on lawns of cells carrying an indicated plasmid. Bars represent mean protection levels obtained from three independent experiments. Error bars show standard errors of the mean. Statistical significance was assessed using Kruskal–Wallis test. ***P < 0.001.