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. 2017 Jul 19;7:5814. doi: 10.1038/s41598-017-06261-5

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© The Author(s) 2017

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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POL elicited TLR6 signaling pathway in immunized C57BL/6 mice. (A) 3 days after the 3^rd vaccination, total RNA from spleens was extracted and qPCR was performed for mRNA expression of TLR6. (B) 3 days after the 3^rd vaccination, the splenocytes were prepared and stained with anti-mouse CD11c-FITC/TLR6-APC mAbs. The CD11c⁺ cells were gated and CD11c⁺TLR6⁺ cells were counted relatively to total CD11c⁺ cells. (C) 3 days after the 3^rd vaccination, the splenocytes were prepared. The cytoplasm proteins or nuclei protein fractions were separated for western blot to examine the levels of MyD88, IRAK1, TRAF6, p-IκB and p-NF-κB. (D) 3 days after the 3^rd vaccination, total RNA from spleens was extracted and qPCR was performed for mRNA expression of IL-1β.