Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Jan 20;12:481. doi: 10.1038/s41467-020-20786-w

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 5 — a Map of the investigated interactions. Since we found NlaIII to be the only restriction enzyme that allowed sufficiently high-resolution analysis of RORCE, NlaIII was finally used to digest the RORCE region. The positions of NlaIII sites (vertical lines) in RORCE used in this 3C analysis are indicated as test sites 1 to 8 (TS1 to TS8). The NlaIII site in RORγt gene promoter was taken as the anchor site (AS), from which possible interactions with other regions of RORCE were quantified. The primers used for 3C-qPCR are indicated by simple black arrows. The negative control site (NCS) is another NlaIII site upstream of RORCE, which does not interact with the AS. b–f Quantitative analysis of a 3C-qPCR assay assessing the RORCE region. The data represent relative interaction frequencies between AS and the other tested sites (TS1 to TS8) in the RORCE region. Relative interaction frequencies were determined by qPCR relative to standard curves as described in the “Methods” section. Data points represent the mean ± SEM for PI-stimulated EL4 (b, n = 4 independent experiments), FACS-purified CD4⁺IL17⁺ Th17 (c, n = 5 independent experiments), Th17-polarized (d, n = 5 independent experiments), Th1-polarized (e, n = 5 independent experiments), and Th2-polarized cells (f, n = 5 independent experiments) from WT mice. Th17 D, Th1 D, and Th2 D represent Th17, Th1, or Th2 differentiation under the appropriate polarization conditions, respectively, and N represents naïve cells. All P-values are based on unpaired two-tailed Student’s t-test (b–d). Experimental mice were between 8 and 12 weeks of age, with no preference to gender and were maintained on a C57BL/6 background. Source data are provided as a Source Data file.