Fig 2: Phenotypic proteins and transcripts expression of MB stem cells models(A) Images of medullospheres of MB stem cells from cell lines: DAOY, D283-Med and D341-Med (× 40 magnification, Bars:100 μm). Expression of CD133 (B), CD15 (C) and NF-M (D) between differentiated cells and MB stem cells by Western blot normalized by β-actin expression. (E) Gene expression of phenotypic transcripts of CD133, CD15 and Sox2 of differentiated cells and MB stem cells normalized by RNA pol II expression. *p < 0.05, **p < 0.01, ***p < 0.001, n = 3.

Fig 3: Immunohistochemical analysis of Nefm expression in the medial prefrontal cortex of MIA-induced mice. (A) Confocal microscopy of Nefm immunofluorescent staining (secondary anti-mouse, green; Alexa Fluor 488); nuclei were stained with DAPI (blue). Treatment with PG inhibited this decrease. (B) Quantitative analysis of Nefm expression revealed a significant decrease of Nefm expression in the MIA group compared with CON. Scale bar, 50 µm. Data are presented as the mean ± standard error of the mean; *P<0.05 vs. CON; #P<0.05 vs. MIA. CON, control group; MIA, maternal induced activation group; Nefm, neurofilament M; PG, Panax ginseng-treated MIA group.

Fig 4: TLR4 deficiency attenuated cognitive impairment and white matter lesions induced by BCAS. (A) The working memory and reference memory of mice were assessed by the 8-arm maze test at 1-month post injury. WT mice suffered from BCAS made much more revisiting errors (P<0.001) and less different arm choices (P<0.001) comparing to the sham-operated mice. TLR4 knockout mice with BCAS made much less revisiting errors (P<0.001) and more different arm choices (P=0.007) comparing to the WT group. No impairment in spatial reference memory was revealed between different groups (P>0.05). Two-way analysis of variance (ANOVA) with repeated analysis, n=9 in each group. (B) White matter lesions were detected by LFB staining in different groups. Scale bar, 200 µm. Dot-plot with median and interquartile range of the severity of white matter lesions in in CCm, CPu, AC, IC, F and OT in histogram. Two-way ANOVA with Dunnett's post-hoc test, **P<0.01 versus WT Sham, #P<0.05, ##P<0.01 versus WT BCAS. n=6 per group. (C) Representative images depicting immunofluorescent labeling of myelin-associated glycoprotein (MAG), myelin basic protein (MBP), and nerve fiber (NF) in the corpus callosum of coronal slices from different groups. Scale bar, 100µm. The expression of MAG, MBP and NF (three major NF subunits based upon their molecular mass: the lowest (NF-L), the middle (NF-M) and the highest (NF-H)) was determined by Western blot in mice from different groups. Quantitative analysis of Western blot results was performed. Two-way ANOVA with Dunnett's post-hoc test, **P˂0.01 versus WT Sham, ##P<0.01 versus WT BCAS. n=8 per group.

Fig 5: Protein expression levels of Nefm, Lasp1, Dpysl2 and Dlg4 in the medial prefrontal cortex of MIA-induced mice. (A) Protein expression levels were determined by western blot analysis; Actb was used as an internal control. Nefm, Lasp1, Dpysl2 and Dlg4 expressions were decreased in the MIA group compared with CON, and treatment with PG inhibited this decrease. (B-E) Quantitative analysis of (B) Dpysl2, (C) Lasp1, (D) Nefm and (E) Dlg4 expression. Data are presented as the mean ± standard error of the mean. *P<0.05 vs. CON; #P<0.05 vs. MIA. Actb, β-actin; CON, control group; Dlg4, discs large MAGUK scaffold protein 4; Dpysl2, dihydropyrimidinase like 2; Lasp1, LIM and SH3 protein; MIA, maternal induced activation group; Nefm, neurofilament M; PG, Panax ginseng-treated MIA group.