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. 2019 Nov 27;9:17662. doi: 10.1038/s41598-019-54198-8

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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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Cardiolipin efficiently reconstitutes NhaA dimers from monomers. Samples of monomeric NhaA suspension (12 µL, 1.7 µg protein, 7.6 µM NhaA) were prepared. One sample with no addition served as a control, and to the other samples different phospholipids were added at the indicated phospholipid/NhaA molar ratios: cardiolipin (CL) or L-a-Phosphatidylglycerol (PG) or L-a-phosphatidyl ethanolamine (PE). The phospholipid-containing samples were sonicated for 10 s in a bath sonicator at 23 °C and incubated for 45 min at 4 °C with slow agitation. Then, 15 µL of native gel sampling buffer was added, and the proteins were resolved on native gels. The experiment was conducted three times with identical results.