Fig. 1

Nedd4 potentiates Th17 cell differentiation in vitro and in vivo. A–C Th1, Th2, and iTreg cell differentiation assays using naïve CD4⁺ T cells from Nedd4^CreER mice pretreated with tamoxifen or corn oil (n = 3 mice per group). D Pathogenic vs. non-pathogenic Th17 cell differentiation assays using naïve CD4⁺ T cells from in Nedd4^CreER mice pretreated with tamoxifen or corn oil (n = 3 mice per group). ***p < 0.001; student t test. E Th1 and Th17 cells in the lamina propria of Nedd4^CreER mice small intestinal tissue pretreated with tamoxifen or corn oil (n = 3 mice per group). **p < 0.01; student t test. F Immunoblot analysis of NEDD4, NEDD4-2, and RORγT in CD4⁺ T cells from MS patients (n = 6) and age- and sex-matched healthy controls (n = 6). *p < 0.05, **p < 0.01; student t test. r = 0.737, Pearson correlation coefficient. The relationship between two variables is generally considered strong when their r value is larger than 0.7. G Expression of NEDD4 and RORγt ex vivo in total CD4⁺ T cells, vs. naïve and memory CD4 + T cells from MS patients and healthy controls (HC) by flow cytometry. Cumulative data from multiple donor samples (10 MS patients and 13 healthy donors). *p < 0.05, **p < 0.01, ***p < 0.001; unpaired student t test. H Flow cytometry evaluating Th17 cell differentiation of human naive CD4⁺ T cells from six healthy blood donors nucleofected with Ctr siRNA or Nedd4 siRNA at the temporal peak of differentiation conditions. **p < 0.01, ***p < 0.001; paired student t test. I NEDD4 knock down efficiency was determined by immunoblot analysis of lysates of T cells transfected with NEDD4 siRNA or control siRNA on day 3