Fig 2: Increased expression of HuR and glial cells in spinal cords of EAE mice. (a) Representative images (low and high magnification) showing immunofluorescence staining for HuR in the spinal cords of control (CTRL), EAE, or anti-HuR ASO-treated EAE mice at 30 days postimmunization. (b) Quantification of the increased HuR immunostaining in spinal cords and significant reduction by anti-HuR ASO. Representative images (low and high magnification) show the immunostaining for GFAP (c), IBA1 (e), and CD11b (f) in the spinal cord of control (CTRL), EAE, or EAE + anti-HuR ASO mice at 30 days postimmunization. Quantification of the GFAP (e), IBA1 (f), and CD11b (h) content showing the increased expression in the EAE section and attenuation by anti-HuR ASO. CTRL, dODN-treated nonimmunized mice; EAE, dODN-treated EAE mice; EAE + ASO, anti-HuR ASO-treated EAE mice. **p < 0.01, ***p < 0.001 versus CTRL; °p < 0.05, °°°p < 0.001 versus EAE (ANOVA). Error bars represent the standard error of the mean. Antibodies are shown on the left. Scale bar = 50 µm (HuR, IBA1, CD11b); 100 µm (GFAP)

Fig 3: MSC transplantation decreased the expression of TLR2 in neurons and suppressed apoptosis in the cerebral cortex of HIBD rats. a Double immunofluorescence staining of NSE or GFAP or Iba1 (green) together with TLR2 nred) in the cerebral cortex of rats of the control, HIBD and MSCs groups. a, d and g. Double immunofluorescence staining of NSE together with TLR2. The white arrows indicate co-localization of NSE and TLR2. b, e and h. Double immunofluorescence staining of GFAP together with TLR2. c, f and i. Double immunofluorescence staining of Iba1 together with TLR2. Scale bar = 50 µm. n = 4 in each group. b TUNEL staining in the rat cerebral cortex of the control, HIBD and MSCs groups. a, d and g. TUNEL-positive cells. b, e and h. DAPI-stained cells. c, f and i. Merged images of TUNEL-positive cells and DAPI-stained cells. The white arrows indicate TUNEL-positive cells that were merged with DAPI-stained cells. Scale bar = 50 µm. c The percentage of TUNEL-positive cells in the three groups of control, HIBD and MSCs. n = 5 in each group. The results are presented as the mean ± SEM. # P < 0.05 vs. the control group; &P < 0.05 vs. the HIBD group

Fig 4: Activation of microglia in the injured area of mice with acute intracerebral hemorrhage. (A) Coronal section of hematoma to observe the bleeding in the brain of mice, n = 6 (Scale bar = 100 µm). (B) Detection of thrombin by immunohistochemistry assay, n = 6. (C–E) Immunofluorescence analysis was conducted to observe the activation of mouse microglia, including the relative activated Iba1+ cells, relative CD86+/Iba1+ cells, iNOS+/Iba1+ cells, CD206+/Iba1+ cells and CD163+/Iba1+ cells, n = 6 (Scale bar = 50 µm). *p < 0.05 vs. sham or ICH 24 h **p < 0.01 vs. sham or CD86/Iba1 (%). ***p < 0.001 vs. sham. ICH, intracerebral hemorrhage.

Fig 5: Impact of age and CR on the microglia marker Iba1 in rat cortex and hippocampus. Representative Western blots and densitometric analysis of Iba1 levels in the cortex (a) and hippocampus (b) normalized to vinculin. 3M (3-month-old rats); 24M (24-month-old rats); ND (normal diet); CR (caloric intake reduced by 40% for the last 6 months of life). Statistical analysis was performed by using one-way ANOVA followed by Tukey’s post hoc test. *** p < 0.001 vs. 3M ND; ## p < 0.01 vs. 24M ND.