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. 2020 May 26;11:2623. doi: 10.1038/s41467-020-16397-0

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© The Author(s) 2020

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. 4 — a Translocation of VpdB. CHO-FcγRII cells were infected with the Legionella strains expressing the indicated VpdB constructs fused to CyaA, and the cAMP production was measured. Shown at the top is protein immunoblotting of CyaA-VpdB expressed in each Legionella strain using anti-CyaA IgG. The expression level of the tested VpdB constructs was similar. Each experiment was biologically duplicated and technically triplicated. b Translocation of SidH. Translocation patterns of SidH and its variants are similar to those of VpdB. Translocation assay and immunoblotting were performed similarly as in (a). c Dependency of LvgA-mediated translocation on IcmSW. Lp01 ΔicmSW and Lp01 ΔicmSWΔlvgA strains exhibited a similar level of attenuation in the transport of VpdB and SidH. d LvgA-dependent translocation of SetA and PieA. Both effectors depended on LvgA to be translocated. CyaA-RalF was used as a control. Each experiment was triplicated, including biological duplication. For immunoblotting, 10 milliOD₆₀₀ units were loaded. Statistical differences compared with the results obtained with Lp01 ΔicmSW were determined by the two-sided t-test for the other strains. The error bars represent mean values ± SD. ***p < 0.001; **p < 0.01; *p < 0.05; n.s., not significant. Exact p-values are provided in Supplementary Fig. 4.