Fig 2: Effects of treatments on differentiation of newly formed cells toward astrocytes in PSD rats at 2, 4, and 8 weeks post-stroke. To observe the differentiation of newly formed cells toward astrocytes, newly formed cells were labeled with BrdU (red) and astrocytes were labeled with GFAP (green) at 2, 4, and 8 weeks. Cell nuclei were labeled with 4',6-diamidino-2-phenylindole (blue). (A) At 2 weeks post-stroke. (B) At 4 weeks post-stroke. (C) At 8 weeks post-stroke. Scale bar = 30 µm. (D) The number of GFAP-positive cells at the observed dentate gyrus zone at 2, 4, and 8 weeks (at 2 weeks: F = 20.579, P < 0.001; at 4 weeks: F = 31.673, P < 0.001, at 8 weeks: F = 13.733, P < 0.001). (E) The number of cells double-positive for GFAP and BrdU at the observed dentate gyrus zone (at 2 weeks: F = 17.182, P < 0.001; at 4 weeks: F = 22.852, P < 0.001; at 8 weeks: F = 6.274, P = 0.001). n = 6; *P < 0.05, *P < 0.01. BrdU, 5-bromo-2-deoxyuridine; PSD, post-stroke depression; FXT, fluoxetine hydrochloride capsules; YNJYP, Yi-nao-jie-yu prescription.

Fig 3: Decitabine treatment increases the expression of p27 in astrocytes.(a) The immunofluorescence staining demonstrates the co-localization of astrocyte marker GFAP (red) with p27 (green) and nuclear marker DAPI (blue) in the peri-infarct area on day 4 after MCAO. Scale bar = 20 µm. (b) Quantitative analysis shows the p27 expression changes with GFAP in each group, n = 6/group, *p < 0.05 by one-way ANOVA plus Tukey’s test.

Fig 4: Pearson's correlation test between the hippocampus expression of GFAP, IBA1, IL-1ß, IL-6, TNF-a, BACE, Aß1-42, p-Tau, the number of rearing in the OFT and the preference index of novel object in the NOR. A The number of rearing was positively correlated to the expression of BDNF. B The preference of novel object was positively correlated to the expression of BDNF. C Expression of Syt-1 was positively correlated to BDNF. D Expression of IBA1 was positively correlated to BDNF. E Expression of IL-1ß was positively correlated to BDNF. F Expression of Aß1-42 was positively correlated to BDNF

Fig 5: Early sex-specific responses to poly I:C in the brain of P8 mice. Increased caspase-3 (A) and caspase-8 activity (B), IRF3 nuclear translocation (D), and INF-ß gene upregulation (F) occur 6 h after poly I:C in the brain of female mice. Increased NF?B (C) and IRF3 (D) nuclear translocation, GFAP expression (E) and INF-ß gene upregulation (F) occur 14 h after poly I:C in the brain of male mice. Data are presented using box and whiskers plots, where boxes are limited by the 75th and 25th percentiles interquartile range (IQR)] and whiskers are limited by the last data point within 1.5 times the IQR from the median (continuous line inside the box). (A–E) White boxes represent NS-treated pups, while gray boxes represent poly I:C-treated pups. (D) Representative immunoblots showing translocation of IRF3 and AP1 from the cytosolic fraction (CF) to the nuclear fraction (NF) in males and females at 6 and 14 h after poly I:C. Top immunoblots show purity of the NF and CF using Lamin B1 and HSP90, respectively. (E) Representative immunoblots showing GFAP expression in cytosolic fractions. (F) Shows the fold change in INF-ß mRNA levels in the brain of male and female mice at 6, 14, 24 h and 7 days after poly I:C relative to NS-treated mice (discontinuous line sitting at 1). °, outlier (between 1.5 and 3 times the IQR); •, extreme (> 3 times the IQR); *p < 0.05.