Fig 2: Impairment of myelin formation in the brain of CpGfapcKO mice.A-D Western blot analysis of GALc and MBP expression levels in the cerebral cortex (A) and hippocampus (C) of Cpflox/flox and CpGfapcKO mice. Quantification of the expression of GALc and MBP in the cerebral cortex (B) and hippocampus (D) was performed. (n = 3). E Immunohistochemistry images of MBP immunostaining in coronal brain sections from Cpflox/flox and CpGfapcKO mice. Scale bar: 1 mm. F: Quantification of MBP area (mm2) in corpus callosum (n = 3). G-L The myelin was examined in the cerebral cortex (G) and hippocampus (H) by TEM, I and J show the corresponding G-ratio in the cerebral cortex (n = 34 from 3 mice); K and L show the G-ratio in the hippocampus (n = 50 from 3 mice for the Cpflox/flox group and n = 48 from 3 mice for the CpGfapcKO group). Scale bar: 1 µm. All the data are presented as the mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001 vs. Cpflox/flox.

Fig 3: Regional expression of MBP and effects of sleep restriction and stress in the frontal cortex, parietal cortex, hippocampus, striatum, thalamus, and cerebellum. Data were normalized to controls for each region. The region × condition interaction was statistically significant (p = 0.008). Bonferroni-corrected post hoc tests showed that sleep restriction resulted in significantly decreased MBP in the striatum relative to controls (p = 0.027). Each point is the normalized value in a single animal. Lines represent means ± SEMs for 5 control, 4 stress, and 5 sleep-restricted mice. *p = 0.05.

Fig 4: Effects of catalpol on myelin protein expression in OLs in EAE mice. Representative immunofluorescence images in the corpus callosum showing MBP (a), GST-pi (b), and NG2 (b) at 20 dpi and 40 dpi in EAE mice. Scale bar: 100 µm in (a); 50 µm in (b). Quantitative analysis of MBP expression (c) (d), GST-pi expression (e), and NG2 expression (f) among groups. Representative western blotting images show the expression of NG2 and Olig2 (g) and GST-pi and MBP (h) at 20 dpi and 40 dpi. Quantitative analysis of relative NG2, Olig2, GST-pi, and MBP levels is done. Protein expression was standardized using Tubulin as a sample loading control, and quantification is presented in each panel. (i) The mRNA expression of Olig2, GST-pi, and MBP at 20 dpi was measured by qRT-PCR and normalized to that of GAPDH. (j) The mRNA expression of Olig2, GST-pi, and MBP at 40 dpi was detected by qRT-PCR and normalized to that of GAPDH. Data are presented as the mean ± SD analyzed by one-way ANOVA with Tukey's post hoc multiple comparison test. #P < 0.05, ##P < 0.01, and ###P < 0.001 versus NC group; *P < 0.05, **P < 0.01, and ***P < 0.001 versus EAE + PBS group.

Fig 5: Myelin protein levels in the cortex in wildtype and St8sia2 -/- mice. Representative Western blots and densitometric analyses of MBP, CNP, MOG, PLP1, and MOBP levels in the cortex in St8sia2+/+ and St8sia2 -/- mice on P15, P30, and P90. The band intensity was normalized to pan-Cadherin (CDH). Each dot represents one animal. The mean for the control is set as 1. The data are expressed as mean ± SD. *P = 0.05; **P = 0.01; ***P = 0.001. [Color figure can be viewed at wileyonlinelibrary.com]