Independent analysis was performed by two blinded observers using a Leica DMRBE microscope equipped with a Cy3 and 4',6-diamidino-2-phenylindole (DAPI) filters.
To detect de- and remyelination in the corpus callosum myelin stained sections for LFB, MBP, MOG, and PLP were scored by three blinded observers using a scale from 0 (complete demyelination) to 3 (normal myelin).
Slides were examined by three independent, blinded observers using a Leica DMIRE2 microscope (Leica DMIRE2, CCD: DFC 420FX, Germany).
As indictated in Materials and methods, stained sections were scored by two blinded observers using a semiquantitative scale from 0 (lowest expression) to 3 (highest level of expression).
Pathology scores were evaluated for each tissue section by three blinded observers using a modified Mankin scoring system, as described [ 27].
After the GPs' performance had been videotaped, it was evaluated by two blinded observers using a checklist to score the presence of key features of the EAP.
Part of the sections were stained with Harris hematoxylin and eosin (HE), and examined microscopically by two blinded observers using a 40× objective lens of a light microscope (Nikon, Tokyo, Japan) connected to a digital camera (Coolpix 990; Nikon).
Disease activity of the CIA was assessed every other day between days 21 and 42 by two blinded observers using a three-point scale for each paw: 0, normal joint; 1, slight inflammation and redness; 2, severe erythema and swelling affecting the entire paw, with inhibition of use; and 3, deformed paw or joint, with ankylosis, joint rigidity, and loss of function.
