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. 2021 Feb 26;11:145. doi: 10.1038/s41398-021-01266-1

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© The Author(s) 2021

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. 3 — Protein extracts obtained from the NAc of male and female P rats (n = 3–4/grp) after administration of 3α,5α-THP (15 mg/kg) or vehicle (45% w/v 2-hydroxypropyl-β-cyclodextrin) were immunoprecipitated (IP) with antibody to MyD88 (A, B) or TRIF (C). Proteins immunoprecipitated with MyD88 antibody were analyzed by immunoblotting (IB) with TLR4 and MyD88 antibodies (A) and TLR7 and MyD88 antibodies (B). Proteins immunoprecipitated with TRIF antibody were analyzed by immunoblotting with TLR4 and TRIF antibodies (C). Normal IgG was used as immunoprecipitation control. MyD88 co-precipitated with TLR4 (A) and TLR7 (B), and TRIF co-precipitated with TLR4 (C) but MyD88 and TRIF did not co-precipitate with normal IgG (A–C). A The levels of TLR4 co-precipitating with MyD88 were significantly reduced by 3α,5α-THP both in males (n = 3/grp) and females (n = 4/grp) (t-test, *p < 0.05). B The levels of TLR7 co-precipitating with MyD88 were also significantly reduced by 3α,5α-THP in females (t-test, *p < 0.05, n = 3/grp). C The levels of TLR4 co-precipitating with TRIF were not altered by 3α,5α-THP treatment both in males (n = 3/grp) and females (n = 3/grp) (t-test, p > 0.05).