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. 2020 Dec 3;10:21095. doi: 10.1038/s41598-020-77858-6

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© The Author(s) 2020

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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Characterization of RORα MKO mice. (A) mRNA expression levels measured by RT-qPCR for Lyz2 and Cre genes in spleen from WT (Rora^+/+ Lyz2^Cre/+) and MKO (Rora^fl/fl Lyz2^Cre/+) mice. (B) PCR analysis of genomic DNA extracted from sorted splenic macrophages of WT and MKO mice. Locations of primer hybridization are represented with filled diamond. Agarose gels were acquired with a Gel Doc XR system (Bio-Rad) and the Image Lab software verion 2.0 build 8 for PC (Bio-Rad, https://www.bio-rad.com/fr-fr/product/image-lab-software?ID=KRE6P5E8Z). Images were cropped for sake of clarity and full-length gels are presented in Supplementary Fig. 6. (C) mRNA expression levels measured by RT-qPCR for the exon 3 of Rora in B cells, T cells, neutrophils, macrophages, monocytes, and dendritic cells (DCs) sorted from the spleen of WT and MKO mice. (D) mRNA expression levels measured by RT-qPCR for the exon 3 of Rora in total bone marrow (BM) cells, in neutrophils purified from BM and in BM-derived macrophages (BMDM) from WT and MKO mice. (E,F) mRNA expression levels measured by RT-qPCR for Rorc (D) and Arntl (E) genes in B cells, T cells, neutrophils, macrophages, monocytes, and DCs sorted from the spleen of WT and MKO mice. (G) mRNA expression levels measured by RT-qPCR for Arntl in BMDM. (H) mRNA expression levels measured by RT-qPCR for the exon 3 of Rora, Arntl, Rorc and Adgre1 in adipose tissue macrophages (ATM) sorted from epididymal adipose tissue of WT and MKO mice. (I) mRNA expression levels measured by RT-qPCR for the exon 3 of Rora in liver, epididymal adipose tissue (epiAT) and skeletal muscle from WT and MKO mice. Data are shown as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 by 2-way ANOVA followed by Sidak’s multiple comparisons test or unpaired t test. All statistical analyses were carried out using GraphPad Prism 8 for Windows (GraphPad Software). n = 8–10 mice per genotype for A & I. n = 5 mice per genotype for C,E & F. n = 3 mice per genotype for D & G. For H, each dot represents a pool of 3–4 mice. NS: Not significant; AU: Arbitrary unit; FC: Fold change. See also Fig. S1.