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. 2018 Mar 9;8:4310. doi: 10.1038/s41598-018-22558-5

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© The Author(s) 2018

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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P. aeruginosa-induced TRPV2 recruitment into lipid rafts at the plasma membrane of macrophages. (A) Confocal images of TRPV2 localization in human primary macrophages after P. aeruginosa treatment (MOI 50, 60 min). Colocalization of TRPV2 staining with the raft marker dye, CTX-B, which recognizes the GM1 ganglioside. Data are representative of three independent experiments corresponding to an average of 63 cells from at least four fields each. Fluorescence profiles corresponding to the line are present below. (B) Human macrophages infected with P. aeruginosa (MOI 50, 60 min) were analyzed by quantitative separation of C + DSM (cytoplasm and detergent-soluble membrane) and N + DRM (nuclei and detergent-resistant membranes) fractions using lysis gradient centrifugation. Both fractions were resolved on reducing gel and analyzed by immunoblotting. Representative blots of four separate experiments are shown. CD71 and flotillin-1 are cell compartment markers for C + DSM and N + DRM respectively. In the histogram, the densitometric analysis of TRPV2 in the N + DRM fraction of the same experiments are shown as mean ± s.e.m. Full-length blots are presented in Supplementary Figure 4. The results were expressed vs. the relative intensity observed in control. Mann-Whitney test: *p < 0.05 vs. control.