Fig 1: BSYSC improved the clinical scores, ameliorated demyelinating lesions, and relieved axonal injury in mice with EAE. (a) The clinical score, the highest average clinical score, and the cumulative clinical score for each group of mice. (b) Fluorescence staining for MBP and Olig2 was performed in the brains and spinal cords of mice. Semiquantitative analysis of MBP fluorescence in the brains and spinal cords of mice was performed with ImageJ. Cell nuclei were stained with DAPI. (c) Fluorescence staining of NF68, NF160, and NF200 was performed in the brains and spinal cords of mice. Semiquantitative analysis of NF68, NF160, and NF200 fluorescence in the brains and spinal cords of mice was performed with ImageJ. Cell nuclei were stained with DAPI, The data are expressed as the mean ± SE (n = 4 in each group at each time point, and each sample was repeated three times). NC: normal control group, EAE: model group, EAE + PA: PA-treated group, and EAE + BSYSC: BSYSC-treated group. *p < 0.05 or **p < 0.01 versus the NC group and #p < 0.05 or ##p < 0.01 versus the EAE group.
Fig 2: Sevoflurane regulates miR-18a-mediated RUNX1/Wnt/ß-catenin signal pathway to suppress NSC proliferation and rat neurodevelopment.A The mRNA expression of RUNX1 under different concentrations of sevoflurane treatment detected by RT-qPCR. B The mRNA expression of ß-catenin under different concentrations of sevoflurane treatment detected by RT-qPCR. C The protein expression of RUNX1 and ß-catenin under different concentrations of sevoflurane treatment detected by Western blot analysis. D The expression of Ki67 protein in NSCs after different concentrations of CWP232228 treatment detected by IHC staining (bar = 25 µm); PI: the nucleus in red fluorescence, Ki67: green fluorescence. E Intuitive and statistical plots of sphere-forming ability of NSCs after sevoflurane, 3 f, miR-18a inhibitor, or CWP232228 treatment (bar = 100 µm). F The effect of sevoflurane, 3 f, miR-18a inhibitor, or CWP232228 treatment on the movement time of rats in the water maze assessed by mirris water maze test. G The effect of sevoflurane, 3 f, miR-18a inhibitor, or CWP232228 treatment on the movement distance of rats in the open field assessed by open field test. H The effect of sevoflurane, 3 f, miR-18a inhibitor, or CWP232228 treatment on the freezing time to cue of rats assessed by conditioned fear test. I The effect of sevoflurane, 3 f, miR-18a inhibitor, or CWP232228 treatment on the freezing time to context of rats assessed by conditioned fear test. J The expression of neural cell marker protein (NF-H/NF-M/NF-L) in cells and tissues detected by Western blot analysis. *p < 0.05 vs. the control/sevoflurane group; #p < 0.05 vs. the sevoflurane + miR-18a inhibitor/sevoflurane + antagomiR-18a group. n = 6. The cell experiment was repeated 3 times independently.
Fig 3: Sevoflurane induces miR-18a expression to affect NSC proliferation and neurodevelopment in rats.A The effect of 4.8% sevoflurane on the NSC proliferation detected by EdU staining (bar = 25 µm) (DAPI: the nucleus in blue fluorescence, EdU staining: red fluorescence). B The effect of 4.8% sevoflurane on the NSC sphere-forming ability (bar = 100 µm). C The efficiency of miR-18a knockdown or overexpression detected by RT-qPCR. D The effect of miR-18a expression on the proportion of Ki67 positive cells under sevoflurane treatment detected by immunofluorescence assay (bar = 25 µm); PI: the nucleus in red fluorescence, Ki67: green fluorescence. E The sphere-forming ability of NSCs after treatment of sevoflurane or miR-18a inhibitor (bar = 100 µm). F The effect of miR-18a expression under sevoflurane treatment on the movement time of rats in the water maze assessed by Morris water maze test; n = 6. G The effect of miR-18a expression under sevoflurane treatment on the movement distance of rats in the open field assessed by open field test; n = 6. H The effect of miR-18a expression on the freezing time to cue of rats under sevoflurane treatment assessed by conditioned fear test; n = 6. I The effect of miR-18a expression on the freezing time to context of rats under sevoflurane treatment assessed by conditioned fear test; n = 6. J The expression of neural cell marker proteins (NF-H/NF-M/NF-L) in cells and tissues after sevoflurane or antagomiR-18a treatment detected by Western blot analysis. *p < 0.05 vs. the control/mimic NC group, #p < 0.05 vs. the inhibitor NC/sevoflurane + inhibitor NC/sevoflurane + antagomiR-NC group. Data were measurement data, expressed as mean ± standard deviation, and analyzed using independent sample t-test. The cell experiment was repeated 3 times independently.
Fig 4: BG45 increased cell viability and the expression of SYT-1 and NF-L. (A) The viability of cells treated with BG45 (0.5, 10, 15, and 25 µM) for 24, 36, and 48 h. (B) Immunofluorescence staining analysis of SYT-1 (red) in primary neurons treated with BG45 (10 µM) for 48 h. (C) Immunofluorescence staining analysis of NF-L (red) in primary neurons treated with BG45 (10 µM) for 48 h. (D) Quantitative SYT-1 analysis. (E) Quantitative NF-L analysis. The values are presented as the mean ± SD from three independent experiments. * p < 0.05 and ** p < 0.01. Scale bar = 50 µm.
Fig 5: Effects of BG45 on the number of dendritic spines, the Aß burden and the expression of synapse-related proteins in APP/PS1 mice. (A) Spine densities of Golgi-Cox-impregnated prefrontal cortex neurons in each group (scale bar = 10 µm) and quantification of spine density. (B) Immunohistochemistry analysis of SYT-1 in each group and quantification of SYT-1 expression. (C) Immunohistochemistry analysis of GAP-43 in each group and quantification of GAP-43 expression. (D) Immunohistochemistry analysis of Ng in each group and quantification of Ng expression. (E) Immunohistochemistry analysis of SV2A in each group and quantification of SV2A expression. (F) Immunohistochemistry analysis of NF-L in each group and quantification of NF-L expression (scale bar = 20 µm). The values are presented as the mean ± SD from three independent experiments. (G) The number of Aß plaques in the prefrontal cortex area of the mice in each group (scale bar = 100 µm) and the quantification of Aß deposition. * p < 0.05, ** p < 0.01, *** p < 0.001.
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