Supplementary Figure 3: Trabid is dispensable in T cells for T cell activation and EAE induction.

Age- and sex-matched wild-type (WT) and T-cKO mice were subjected to MOG_35-55-induced EAE. (a) Flow cytometry analysis of the CD45⁺ immune cells (CD4⁺ and CD8⁺ T cells and CD11b⁺ monocytes) infiltrated into the CNS (brain and spinal cord) (n = 3, day 14 post-immunization), presented as a representative plot (left) and summary graph (right). (b) Flow cytometry analysis of the IFN-γ-producing T_H1 and IL-17A-producing T_H17 cells in the CNS (percent of CD4⁺CD45⁺ cells) and draining lymph nodes (percent of CD4⁺ T cells) (n = 3, day 14 post-immunization). Data are presented as a representative plot (left, CNS only) and summary graph of absolute cell numbers (right). (c) ELISA analysis of the indicated cytokines in the supernatants of cultures of splenocytes isolated from WT and T-cKO EAE mice (day 14 after immunization) and restimulated in vitro with MOG peptide (20 μg/ml for 48 h). (d) Flow cytometry analysis of the frequency of T_H1, T_H17, and T_REG cells in wild-type and T-cKO naive CD4⁺ T cells, stimulated for 4 days with plate-bound anti-CD3 and anti-CD28 antibodies under T_H0, T_H1, T_H17, or T_REG conditions. (e) Flow cytometry analysis of cell proliferation, based on CFSE dilution, of naïve T cells stimulated with plate-bound anti-CD3 (5 μg/mL) and anti-CD28 (1 μg/mL) for 48 h. (f) ELISA of IFN-γ and IL-2 using supernatants of the cell cultures described in e. Data are representative of three independent experiments. Error bars are mean ± SEM values.