Fig. 4: RNF213 interacts with FOXO1 and is critical to regulate the translocation of FOXO1 to the nucleus.
A Schema of IP-MS approach to identify RNF213-interacting proteins in Treg cells. B Volcano plot of RNF213-interacting proteins by immunoprecipitation–mass spectrometry (IP-MS) analysis. Red indicates significantly enriched proteins (log2(fold change) >1; t-test adjusted P < 0.05). C The list of Ube2d2a, UBA7, SOCS3, FOXO1, TRAF1 and MEX3A identified by IP-MS analysis. D Immunoprecipitation (IP) and immunoblot (IB) analysis of Treg cells isolated from WT or Rnf213Tg mice. E IP and IB analysis of HEK293 cells transfected with the indicated plasmids for 24 h. F Luciferase activity of HEK293T cells transfected with a luciferase reporter driven by the Foxp3 promoter and expressing vector alone or various combinations (horizontal axis) of FOXO1 and RNF213. G Treg cells were double-stained with anti-FOXO1 (red) Abs and DAPI (nucleus, blue) and then observed by fluorescence microscopy. Scale bars: 50 μm. Quantification of fluorescent digital imaging analysis of FOXO1 expression in the cytosol and the nucleus depicted as the percentage expression levels are presented in the right panel (n = 3). Data shown are the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001. P values were calculated using a two-sided, unpaired Student’s t-test in (F). N = 3 (E) repeats from three independent experiments. Source data are provided as a Source Data file.
