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. 2019 Sep 11;9:13146. doi: 10.1038/s41598-019-49543-w

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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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DMBT1 glycosylation is involved in inhibition of P. aeruginosa twitching motility. Effect of DMBT1 digested by deglycosylation enzymes on (A) twitching velocity of P. aeruginosa PAO1 after 4 h, and (B) colony size of P. aeruginosa PAO1 after 4 h and 24 h. Data are shown as the mean ± SEM per sample from three independent experiments. Significance was determined using one-way ANOVA (panel A) or two-way ANOVA (panel B) each with Tukey’s multiple comparisons, ****P < 0.0001; ***P < 0.001; *P < 0.05; ns, not significant. (C) Dot-immunoblot assay with anti-PilA antibody shows continued binding of PAO1 pili to purified DMBT1 (400 ng) after deglycosylation digestion. Pili extracts from PAO1 (S-PAO1), extracts from the pilA mutant (S-PAO1-pilA::Tn), both diluted 1 in 500 in PBS were used as controls, along with PBS alone. (D) Quantification of dot-intensity using ImageJ from the dot-immunoblot assay shown in panel C. Data are shown as the mean ± SEM of duplicate measurements from each sample. Significance determined used Kruskal-Wallis test with Dunn’s multiple comparisons, *P < 0.01; ns, not significant.