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. 2017 Jul 25;7:6349. doi: 10.1038/s41598-017-06484-6

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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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TLR4 deficiency reduces pro-inflammatory chemokines and cytokines expression and classically activated M1 macrophage. Fifty micrograms of protein from each group were separated on SDS-PAGE and incubated with appropriate antibodies overnight. (A) Representative cropped immunoblot images for MCP-1, MIP-2, CD68, a marker for M1 type macrophage, TNF α and IL-6 showing reduced expression in TLR4 deficiency mice in response to Ang-II treatment, (B,C) Data normalized to to β-actin and presented as mean ± SEM, (D) mRNA fold change for MCP-1 and MIP-2 assessed by real-time PCR. Results are expressed as fold change relative to control mice (C3HeOuJ + Saline). n = 5/group, tested by Kruskal-Wallis test and Mann-Whitney rank sum test. *p < 0.05 vs. C3HeJ + saline, ^†p < 0.05 vs. C3HeOuJ + Ang-II, ^‡p < 0.05 vs. C3HeOuJ + saline. C3HeOuJ: normal TLR4, C3HeJ: TLR4 deficiency.