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. 2021 Jul 12;11:14319. doi: 10.1038/s41598-021-93590-1

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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 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/.

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Isolation and validation of microglia-specific mRNA from Cx3cr1^CreER: Rpl22^HA mice. (A) Overview of the experimental strategy of generation of Cx3cr1^CreER: Rpl22^HA mice and isolating RNA. Black ribosomes in the cartoon represent endogenous wt ribosomes, whereas yellow/red are HA tagged transgenic Rpl22 containing ribosomes. (B) Representative images from the amygdala of Cx3cr1^CreER: Rpl22^HA mice validating the specificity of HA expression in microglia. CX3CR1 + cells carry GFP and an antibody against GFP was used to label CX3CR1 + cells (green) and anti-HA was used to label Rpl22^HA expressing cells. The merged imaged shows that HA and GFP are co-localized. (C) Bar graph shows that RNA is significantly enriched in the immunoprecipitated RNA. *indicates p < 0.05 (Tukey’s multiple comparison test). The enrichment in the microglial IP RNA was compared to the total RNA before IP. Three animals per group were used in the experiments described in (B,C).