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. 2021 Nov 1;12:6270. doi: 10.1038/s41467-021-26545-9

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© The Author(s) 2021

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–c To study the heme stealing properties of HphA, 2.5 µM purified WT and H43A/H106A HphA was added to RPMI media in combination with 50 nM hemoglobin (Hb). Growth of hsmA150::Tn26 (a), hphA112::Tn26 (b), and hphR109::Tn26 (c) was measured at 37 °C. d–f OD₆₀₀ at the 12 h timepoint for hsmA150::Tn26 (d), hphA112::Tn26 (e), and hphR109::Tn26 (f). Note: the same WT data was used in all panels for ease of comparison between WT and each individual mutant. All values plotted represent the mean ± standard error of the mean from three separate experiments, and results analyzed by one-way ANOVA followed by Tukey’s test. ns P ≥ 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Adjusted P values are as follows: mutant + Hb vs. WT + Hb P = 0.0043 (d), P = 0.0029 (e), P = 0.0002 (f); mutant + Hb + apo vs. WT + Hb P = 0.2717 (d), P = 0.2331 (e), P = 0.0003; mutant + Hb + H43A/H106A vs. WT + Hb P = 0.0076 (d), P = 0.0022 (e), P < 0.0001 (f); mutant + Hb + apo vs. mutant + Hb P = 0.0005 (d), P = 0.0434 (e), P = 0.9954 (f); mutant + Hb + H43A/H106A vs. mutant + Hb P = 0.9654 (d), P = 0.9962 (e), P = 0.3137 (f); mutant + Hb + H43A/H106A vs. mutant + Hb + apo P = 0.0008 (d), P = 0.0322 (e), P = 0.2337 (f).