Up-regulation of GFAP over the course of MOG-EAE.

<p>(A) Western-Blot analysis of spinal cord protein preparation at the early chronic (day 35 p.i.) and late chronic phase of the disease (day 60 p.i.). Each lane represents the GFAP expression of one single mouse, β-actin serves as a loading control. (B) Quantification of optical densities of the GFAP labeling as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007624#pone-0007624-g002" target="_blank">figure 2A</a>. There was a clear increase in GFAP expression over the course of MOG-EAE (p<0.05 on day 60 p.i.). (C) Immunohistochemistry for GFAP in naïve C57BL/6 mice without EAE (left) and on day 35 p.i. (middle) and 60 p.i. (right). Representative images from spinal cord cross sections are shown, arrows indicate GFAP positive astrocytes which appear elongated in naïve mice and swollen in the chronic phases of MOG-EAE. Bar represents 20 µm.</p

Detection of GFAP in CSF samples of secondary progressive MS patients.

<p>(A) Boxplot analysis of a group of unselected MS patients in comparison to control patients without neurological disease. GFAP levels in the CSF are increased in the multiple sclerosis (MS) group (p<0.05). (B) Boxplot analysis comparing secondary progressive MS (SP-MS) patients, relapsing remitting MS (RR-MS) patients and controls. GFAP levels in the CSF are exclusively increased in the secondary progressive MS group.</p

Statistical evaluation of regulated proteins in CNTF −/− mice.

<p>2D DIGE analysis of the average spot volume increase ratio of proteins shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007624#pone-0007624-g004" target="_blank">Fig. 4</a>. Using the DeCyder's Biological Variation Analysis module, a paired Student's t-test yielded a p-value within the 99th percentile confidence level. Mean value crosses are connected; IS  =  Internal Standard.</p

Proteome analysis of CNTF knockout mice.

<p>2D-DIGE gel of spinal cord proteins from CNTF knock-out mice with EAE, labeled with Cy3 (shown in green) versus wild type mice with EAE labeled with Cy5 (shown in red). Selected proteins identified by Mass Spectrometry are indicated with roman indices (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007624#pone-0007624-g005" target="_blank">Fig. 5</a>). Proteins were separated on 24 cm IPG Strip pH 3–10 non-linear.</p

Proteome analysis of chronic MOG-EAE.

<p>2D-DIGE gel of brain proteins from wild type mice with EAE, labeled with Cy3 (shown in red) versus healthy control (without EAE) labeled with Cy5 (shown in green). Images of gel region containing the glial fibrillary acidic protein (GFAP; identified by Mass Spectrometry) is shown in detail for the respective CyDye. IS  =  Internal Standard (Mixture of Samples). Proteins were separated on 24 cm IPG Strip pH 3–10 non-linear. CRNP =  Calreticulin Precursor; PDI = Protein Disulfid Isomerase Precursor.</p

CNTF deficiency leads to destructive EAE lesions with enhanced apoptosis.

<p>EAE lesions in CTNF −/− mice (left) are destructive with vacuolar degeneration (asterisks) and multiple apoptotic cells with pycnotic nuclei (depicted by arrows). These changes are not present in EAE lesions from wild-type control mice (right side). Representative cervical spinal cord cross sections from a CNTF −/− knockout mouse and a wild-type CNTF +/+ control mouse on day 35 p.i. of MOG-EAE are shown. Bar  = 50 µm.</p