Fig. 6. TRAF2 deletion enhances apoptosis of TNF-stimulated OTUD7b-competent BMDC and protects mice from ECM.

Traf2 was deleted in BMDC using CRISPR/Cas9. A TRAF2-sufficient and -deficient BMDC were either stimulated with 50 ng/mL of TNF for 6 h or left untreated (0 h). Cells were harvested at the respective time points and stained for the indicated proteins by WB. B–D TRAF2-sufficient and -deficient BMDC (1×10⁶ cells per recipient) were i.v. adoptively transferred into CD11c-Cre Otud7b^fl/fl mice followed by i.p. injection with 1×10⁶ PbA-infected RBCs. B Mice were monitored once per day until day 7 p.i. for neurological signs of ECM according to the RMCBS scale (Mann–Whitney U test, n = 10 per group, *p < 0.05). C Peripheral blood parasitemia was determined daily during the course of infection using Giemsa-stained thin blood smears. Data represent mean values + SEM (n = 5 per group, Student’s t-test with *p < 0.05). D Representative images of Evans blue assay performed at day 7 p.i. Mice were injected i.v. with Evans blue dye and perfused with 0.9% saline 1 h postinjection. Brains were isolated and photographed. Representative images of one of two independent experiments with 5 mice per group each are shown.