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. 2019 Nov 28;9:17851. doi: 10.1038/s41598-019-54229-4

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© The Author(s) 2019

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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Fragmentation analysis and AraQ/MalR1 binding. Panel (1) depicts the promoter region of pyk (Bbr_0757) used for fragmentation analysis. −35 and −10 sites are indicated in red and the ATG start codon is indicated in green. The TFBS is indicated in bold. The fragment including the TFBS (Frag. 1) is underlined in blue, the fragment excluding the TFBS (Frag. 2) is underlined in yellow, while the full fragment is referred to as IR. Panel (2) EMSA to investigate AraQ and MalR1 abilities to bind to Bbr_0757 promoter region fragments (IR, Frag 1 and Frag 2). All reactions contain 0.5 nM Ird labelled DNA and 150 nM of either AraQ or MalR1 protein, while negative reactions contain 0 nM protein. Uncropped gel image available in Supplemental Fig. S5. Panel (3), (A) Illustrates consensus AraQ/MalR1 binding motif across the Bifidobacteriaceae family (B) Depicts the binding motif of AraQ/MalR1 in B. breve. (C) Depicts the AraQ/MalR1 binding motif for core members of the regulon.