Fig. 4
From: Loss of the molecular clock in myeloid cells exacerbates T cell-mediated CNS autoimmune disease
Enhanced Th1 and Th17 responses in the CNS of Bmal1 Mye−/− mice with EAE. a–e Bmal1 Myeloid+/+ or Bmal1 Myeloid−/− mice were immunized with myelin oligodendrocyte glycoprotein (MOG35–55) + complete Freund’s adjuvant (CFA) on d 0 and with pertussis toxin (PT) (125 ng/mouse) on d 0 and d 2. a After 14 d draining lymph node (LN) cells were re-stimulated in the presence and absence of MOG35–55 antigen (25 and 50 µg/ml) or with medium for 72 h. Supernatants were removed and tested for IL-17, GM-CSF, IFN-γ and IL-10. Statistics by one way ANOVA with Tukey’s post-test of triplicate assay of 6 mice per group. b–e 10 d post immunization brain and spinal cords were isolated and surface stained for CD45, CD3, CD8α and CD4 and intracellularly for IL-17 and IFN-γ. b Cells were gated on live CD45+ cells in the brain (n = 5–6). c Cells were gated on live CD45+ cells spinal cord (n = 5–6). d Cells were gated on live CD45+CD3+ cells in the brain (n = 5–6). e Cells were gated on live CD45+ cells in the brain (n = 5–6). Statistics were performed by Mann–Whitney U test (b–e). All data presented as means±standard error of the mean (SEM). *p < 0.05; **p < 0.01; ***p < 0.001
