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. 2023 Sep 7;20:204. doi: 10.1186/s12974-023-02889-5

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Microglial response caused by high-dose of estrogen (hE2) via ERα. A Representative images of microglia (Iba1-labeled) and estrogen receptors (Erα, ERβ, and GPER) co-located in mouse brain and BV2 cells by immunofluorescence. Bar = 15 μm. The siRNAs are used to suppress different estrogen receptors (ERs) in BV2 cells. Negative siRNA is used as the negative control. The effects of hE2 on CD86 and Iba1 are subsequently analyzed. B Representative images of microglia (Iba1-labeled) and M1 polarization markers (CD86) co-located by immunofluorescence. C qPCR analysis of ER proteins in BV2 cells transfected with different siRNAs to suppress the corresponding ER. D Quantification of the mean fluorescence intensity per cell with the indicated number of cells. E The siRNAs are used to suppress different estrogen receptors (ERs) in primary microglia. Results of qPCR analysis of ER proteins in primary microglia transfected with different siRNAs to suppress the corresponding ER. Representative band (F) and quantification (G) of protein expression in primary microglia. Students t-test is performed to determine the significant difference based on P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***), respectively, in comparison to the control group as indicated by black asterisks or “ns” (i.e., no statistical significance). Data are presented as mean ± standard deviation (SD)