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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. 1 — a MS spectrum of peptide ¹⁴⁸VQEAASQGLK¹⁵⁷ corresponding to Raftlin1 (UniProt accession No: Q6A0D4). Observed b- and y-ions are indicated. b The lysates from CD4⁺ T cells isolated from colonic lamina propria of C. rodentium infected WT mice were pulled-down with purified recombinant GST or GST-RORγt followed by Immunoassays with anti-Raftlin1 and anti-GST antibody. c GEO dataset (GSE130302) was analyzed (n = 2 C. rodentium-infected and n = 3 SFB-colonized colon) and presented as the MA plot showing the change in expression (log2 fold change) and counts per million (log CPM values); p < 0.05, FDR < 0.05. d Rftn1 mRNA expression was analyzed by real-time PCR using CD4⁺ T cells isolated from colonic lamina propria; n = 6 mice per group. Data are presented as mean value ± SD, p = 0.00010 (CR), p = 0.37 (SFB). e Immunoassays of Raftlin1 protein in the lysates of CD4⁺ T cells. f the lysates of CD4⁺ T cells isolated from colonic lamina propria of C. rodentium infected mice were subjected to immunoprecipitation with anti-Raftlin1 or anti-RORγt antibody and immunoblot analysis with anti-RORγt or anti-Raftlin1 antibody. g Sequence alignment of Raftlin1; box indicates the conserved LLNSL motif. h Putative protein-protein interface from the docking model is shown in color gray for RORγt and Salmon for Raftlin1. Helix 12 (labeled) and AF2 (green) domains of RORγt are involved in non-covalent interaction in the modeled RORγt-Raftlin1 complex. LLNSL region of Raftlin1 is shown in color yellow. i Lysates from HEK293T cells transfected with various combinations of plasmids encoding Flag-RORγt and either Myc-Raftlin1 or Myc-Raftlin1^ΔLLNSL, followed by immunoprecipitation with anti-Flag antibody and immunoblot analysis with anti-Myc or anti-Flag antibody. Immunoblots in (b, e, f, i) are from one experiment representative of three independent experiments with similar results. Statistical significance was determined by unpaired Student’s t test (two-tailed) in (c, d) with p < 0.05 considered statistically significant. ****p < 0.0001; ns- not significant. Source data are provided as a Source data file.