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. 2019 Aug 22;9:12252. doi: 10.1038/s41598-019-48697-x

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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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Electrophoretic protein profiles of LVS variants or mutants grown as a biofilm or are planktonic cells from the biofilm broth supernatant. Bacterial strains were grown as a biofilm in CDMB for 25 days, the medium above the biofilm was removed, and the planktonic cells harvested by centrifugation. Planktonic cells in the medium and cells in the biofilm were extracted with 1 M urea. The protein profiles were resolved by SDS-PAGE on 4-12% gels and stained with silver. Strains: L, LVS; P, LVS_P10; LM, LVSΔ1423-22; WM, WbtI_G191VΔ1423-22; M, Molecular size standards. The figure is a compilation of four separate gels containing samples prepared during the same experiment (left two are from planktonic phase cells and right two are from biofilm cells), which have been cropped to present the most relevant information (original shown in Supplementary Fig. S3). Precipitates of the broth medium are not shown. Lanes of WbtI_G191V (W) have been deleted from the planktonic cells (far left) and from the biofilm cells (far right) due to lack of differences with WbtI_G191VΔ1423-22 (WM). Repeated subculture in CDMB had little effect on protein composition, but loss of surface glycoses, particularly O-Ag, or growth as a biofilm substantially altered the protein profile.