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. 2020 Mar 5;11:1213. doi: 10.1038/s41467-020-14941-6

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Relative normalised fluorescence of C. necator carrying the various inducible system–reporter constructs in response to different concentrations of their corresponding primary inducers. Measurements were taken 80 min after the inducer had been extracellularly added. The dose–responses were fit using a Hill function (see Methods section). The maximum level of reporter output b_max was set to 100% (except in case of the GABA-inducible system where the absolute normalised fluorescence corresponding to the highest GABA concentration tested was set to 100%). The inducer concentration that mediates half-maximal reporter output K_m is indicated by a dotted line. Data are mean ± SD, n = 3, non-linear regression. Source data are provided as a Source Data file. b Evaluation of induction homogeneity by flow cytometry. The fluorescence intensity of 100,000 individual cells was determined for each inducible system–reporter construct 2 h after extracellular addition of inducer. Uninduced cells (grey) are compared to cultures supplemented with their cognate effector at final concentrations corresponding to 50% (orange) and 95% (blue; 90% in case of the phenylglyoxylate-inducible system) of the maximum level of reporter output b_max. The tartrate- and sulfonatoacetate-inducible systems were only evaluated for induction homogeneity at 50% of b_max due to solubility limits and inducer toxicity, respectively. Since b_max could not be calculated for the GABA-inducible system, induction homogeneity was determined using a final concentration of 250 mM (purple).