T cell–specific PIKFYVE deletion does not alter homeostatic development. (A–C) Cells were isolated from the (A) thymus, (B) spleen, or (C) lymph nodes from PIKFYVE^f/f or CD4^CrePIKFYVE^f/f mice under homeostatic conditions. Cells were analyzed by flow cytometry for viability, CD3, CD4, CD44, or CD62L. Data represent mean ± SEM, n = 3. P values were calculated with a Student’s t test. (D and E) The frequency and number of Tregs were analyzed by flow cytometry in the (D) thymus and (E) spleen. (F) Schematic model with the experimental design of EAE is depicted. C57BL/6 mice (PIKFYVE^f/f or CD4^CrePIKFYVE^f/f) were immunized with MOG_35–55 peptide in CFA and time points that were included in the analysis are highlighted (Day 7, 15, or 30). (G) PIKFYVE^f/f or IL17^crePIKFYVE^f/f mice were immunized with 200 µg of MOG_35–55 peptide containing CFA to induce EAE. On days 0 and 2, mice received 200 ng/mouse of pertussis toxin. EAE clinical score was measured daily (N = 5).