Fig. 6. DeepDTnet-predicted topotecan (TPT) reverses experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. (A) An illustration of induction and treatment of EAE. (B) Mean clinical scores of EAE in vehicle- or TPT-treated group (n = 10/group). TPT (10 mg kg−1) or vehicle is intraperitoneal administered on day 11 after immunization every four days. Data are presented as the mean ± SEM of eight mice per group. Student's t-test is revealed, *P < 0.05, **P < 0.01. (C) The body weight of mice in vehicle- or TPT-treated group. Student's t-test is revealed, *P < 0.05. (D) Section of spinal cord tissue is prepared on day 20 post immunization and subjected to hematoxylin and eosin (H&E) staining and Luxol fast blue (LFB) staining. (E) In vivo imaging of myelination using myelin-binding dye, 3,3-diethylthiatricarbocyanine iodide (DBT) on day 20 after immunization. DBT dye readily enters the brain and specifically binds to myelinated fibers. (F) In vivo imaging of the blood–brain barrier integrity using Cy5.5-BSA on day 20 after immunization. Cy5.5-BSA uptake in the brain when the BBB (blood–brain barrier) integrity is disrupted. (G) ELISA analysis of IL-17 production of spinal cords and brain from vehicle- or TPT-treated EAE mice on day 20 after immunization. Data are presented as the mean ± SEM. Student's t-test is revealed, **P < 0.01. (H and I) Concentration of T0901317 in mice brain samples (H) and plasma (I). T0901317 (ref. 56), an orthosteric ligand of ROR-γt, was used as the tracer for assessing target occupancy of TPT in the mouse model. Student's t-test was performed and sterile water was used as vehicle.
