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. 2017 Aug 21;7:8363. doi: 10.1038/s41598-017-08743-y

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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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The ELISA assay of the interaction between TLR2 protein and LipL32 variants. (A) The interaction of TLR2 protein and LipL32 truncated variants. The protein concentration of LipL32 and TLR2 protein were 1 µM. BSA and Pam₃CSK₄ were served as negative and positive controls. WT + PK indicated the LipL32WT pretreated with proteinase K for ELISA assay. (B) The interaction of TLR2 protein and LipL32 point mutation variants. (C) The dose dependent ELISA of the interaction between LipL32ΔCenα3 variant and TLR2 protein. ○, LipL32WT in the presence of 1 µM Ca²⁺; ●, LipL32WT in the presence of 1 µM EGTA; ▼, LipL32ΔCenα3 in the presence of 1 µM Ca²⁺; △, LipL32ΔCenα3 in the presence of 1 µM EGTA. (D) The dose dependent ELISA of the interaction between LipL32 point mutation variants and TLR2. ○, D195A variant in the presence of 1 µM Ca²⁺; ●, D195A variant in the presence of 1 µM EGTA; ▼, D196A variant in the presence of 1 µM Ca²⁺; Δ, D196A in the presence of 1 µM EGTA. *p < 0.05; **p < 0.01; ***p < 0.001.