Fig 2: Immunofluorescence (IF) for GFAP representing astrogliosis in the frontal cortex of fetal baboons from early and mid-gestation ZIKV inoculated dams compared to the control fetal brains.In fetal frontal cortices from both early and mid-gestation ZIKV inoculated dams, significant increase in GFAP+ cell population was seen. GFAP immunoreactive cells also visually appeared to have increased GFAP staining intensity and morphological changes such as longer and more ramified cellular processes compared to control brains (arrow and inset). (Scale bar = 50µm; Error bars depict SD, Dunnett’s multiple comparison test was done for statistical analysis, p<0.05 considered significant).

Fig 3: PGM aggrandized glial cell activation in the hippocampal regions of APP/PS1 mice. (A) Fluorescent GFAP (red)/DAPI (blue) colocalization in the DG regions of the hippocampus in APP/PS1 mice. Scale bar = 100 µm. (B) Fluorescent GFAP (red)/DAPI (blue) colocalization in the DG regions of the hippocampus in WT mice. Scale bar = 100 µm. (C) Fluorescent Iba1 (red)/DAPI (blue) colocalization in the DG regions of the hippocampus in APP/PS1 mice. Scale bar = 100 µm. (D) Fluorescent Iba1 (red)/DAPI (blue) colocalization in the DG regions of the hippocampus in WT mice. Scale bar = 100 µm. (E) Quantification of GFAP-positive cells in each APP/PS1 mice group. n = 5–8 per group, three sections per animal. (F) Quantification of GFAP-positive cells in each WT mice group. n = 5–8 per group, three sections per animal. (G) Quantification of Iba1-positive cells in each APP/PS1 mice group. n = 5–8 per group, three sections per animal. (H) Quantification of Iba1-positive cells in each WT mice group, n = 5–8 per group, three sections per animal. *P < 0.05 vs. APP/PS1-saline, **P < 0.01 vs. APP/PS1-saline.

Fig 4: The influence of LPS on glia cells among the PAD4 KO and wildtype mice. (A) Representative immunofluorescence of Iba-1 in the hippocampus. (B) Column graphs representing Iba-1 expression. (C) Representative immunofluorescence of GFAP in the hippocampus. (D) Column graphs representing GFAP expression. Data were expressed mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001. bar = 100 um; WT Saline, WT LPS, PAD4 KO Saline: n = 3; PAD4 KO LPS: n = 4.

Fig 5: Generation of repair Schwann cells.(a) IF images of sciatic nerve cross sections from WT and PrnpZH3/ZH3 mice at 5 and 10 d.p.c. showed upregulation of c-Jun. Nuclear c-Jun expression was faint in the uninjured sciatic nerve of a WT mouse. Channels for c-Jun and DAPI are shown separately. Scale bar 50 µM. (b) Quantification of c-Jun positive nuclei revealed no significant differences between WT and PrnpZH3/ZH3 mice at 5, 16 and 30 d.p.c, nor between uninjured nerves from WT and PrnpZH3/ZH3 mice (two-way ANOVA, Sidak’s multiple comparisons test, p > 0.05). (c) Western blot of lysates from crushed and uninjured (contralateral) sciatic nerves at 5, 10 and 16 d.p.c. showed a pronounced upregulation of GFAP and total c-Jun protein levels both in WT and PrnpZH3/ZH3 mice. Also, a slight upregulation of actin was detected in crushed nerves. (d) For quantification, GFAP and c-Jun were normalized to actin and expressed as fold change to the average level of crushed WT nerves. No statistically significant difference was detected between WT and PrnpZH3/ZH3 mice at the investigated time points (two-way ANOVA, Sidak’s multiple comparisons test, p > 0.05). (e) Western blot detecting low GFAP levels in uninjured sciatic nerves from WT (n = 6) and PrnpZH3/ZH3 mice (n = 6) sacrificed at 5, 16 and 30 d.p.c. GFAP levels were higher in PrnpZH3/ZH3 sciatic nerves when compared to WT nerves. Blotting with POM2, a monoclonal anti-PrP antibody, detected PrP in WT but not in PrnpZH3/ZH3 sciatic nerves, confirming the absence of PrP in PrnpZH3/ZH3 mice.