*5.2. Experimental Design*

Mice were handled according to Figure 9. Briefly, EAE was induced in female C57BL/6J mice. Pain sensitivity was evaluated before (day 0) and on the fourth, fifth, sixth, eighth, and 10th days (as Section 5.4) after immunization (as Section 5.3). Clinical signs were daily evaluated. CRO was administered in a single dose (on the 5th day after immunization–1st protocol) or in five repeated doses from the 12th day (one daily dose for five consecutive days–2nd protocol). At the 17th (peak of disease) and 28th (remission phase) days after immunization, the spinal cord was collected for inflammatory infiltrate determination (as Section 5.6), glial cells immunoreactivity evaluation (as Section 5.7), cytokines release measurement (as Section 5.8), and nerve growth factor expression (as Section 5.9). At the same periods, the sciatic nerve was removed for myelin thickness determination (as Section 5.10).

**Figure 9.** Experimental design for the evaluation of CRO effect on EAE development.
