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. 2023 Aug 17;14:4972. doi: 10.1038/s41467-023-40622-1

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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/.

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Fig. 2 — a Luciferase assay of lysates of Jurkat T cells transfected with various plasmid combinations (below plot) of the IL-17 promoter-driven luciferase plasmid (pGL4-IL17pr) and the plasmid encoding Flag-RORγt along with wild-type Myc-Raftlin1 or Myc-Raftlin1^ΔLLNSL; results are presented in relative luciferase unit (RLU); n = 3 biological replicates, p = 0.0009, p = 0.002. b Real-time PCR and ELISA were performed to check the expression of IL-17 in Raftlin1 knockdown lysates of CD4⁺ T cells isolated from colonic lamina propria of C. rodentium infected WT mice; n = 3 biological replicates, p = 0.0002 (Il-17a), p = 0.001 (IL-17a). Immunoblot showing knockdown of Raftlin1. c Immunoblot analysis for Raftlin1 in the cytoplasmic and nuclear fraction of CD4⁺ T cells isolated from colonic lamina propria. d Representative images from proximity ligation assay (PLA) showing the interaction between RORγt and Raftlin1. The image is representative of three independent experiments. Scale bars, 5 μm. e Binding of RORγt-Raftlin1 to the IL-17 promoter sites was assessed by ChIP, and re-ChIP analysis using a DNA-protein complex using a specific antibody. f GEO dataset (GSE59071) was analyzed (n = 11 healthy control and n = 74 active UC patients samples), and the normalized expression value of RFTN1 is as shown above, p = 2 × 10⁻⁹. g Expression of IL-17A mRNA and RFTN1 mRNA was analyzed by real-time PCR in active UC and control samples (n = 10 healthy control and n = 11 active UC patients samples). Relative IL-17A mRNA was plotted against relative RFTN1 mRNA; p = 0.0130, Pearson correlation coefficient r = 0.5322. Data in (a–c, e) are from one experiment representing three independent experiments with similar results. Statistical significance was determined by unpaired Student’s t test (two-tailed) in (a, b, f, g) with p < 0.05 considered statistically significant. **p < 0.01; ***p < 0.001; ****p < 0.0001, error bars are mean ± SD. The statistics (g) were measured by the Pearson correlation coefficient. Source data are provided as a Source data file.