Figure 7.

A lack of RGC-32 affects the number of CTGF-positive astrocytes in spinal cords from EAE mice but not in control mice. Spinal cords were harvested from control mice (day 0) and from mice with EAE at day 14 and double-stained with anti-CTGF (red) and anti-GFAP (blue) antibody. At day 0, CTGF⁺ astrocytes are barely detected in both groups (A, B, arrows), but during EAE they become abundant in WT mice (C, arrows), as compared to RGC-32 KO mice, where CTGF co-localization remains at a low level (D, arrows). Controls (CTR) for the immunoperoxidase and alkaline phosphatase reactions were negative (E). Original magnification: x40. Scale bars: 20 µm. CTGF/GFAP-double-positive cells were manually counted in white matter areas corresponding to 20x-magnified spinal cord sections, and the mean value per area was compared between WT and RGC-32 KO mice, and between day 0 and day 14. No difference was detected between WT and RGC-32 KO mice at day 0. A significantly higher number of astrocytes with CTGF co-localization was observed in WT mice than in RGC-32 KO mice at day 14 (F). Only WT mice showed a significant increase in CTGF⁺ astrocytes at day 14 as compared to day 0 (F). Results in (F) are expressed as mean ± SEM (Day 0: n = 6 areas per 2 mice in WT; n = 8 areas per 2 mice in RGC-32 KO. Day 14: n = 7 areas per 2 mice in WT; n = 8 areas per 2 mice in RGC-32 KO). **** = p<0.0001; ns = not statistically significant.