NeuN Antibody - #DF6145

Figure 2 |Immunofluorescence staining of Beclin 1, NeuN and GFAP in the dorsal column along the epicenter at 24 hours after SCI (fluorescence microscope). Double staining of Beclin 1/NeuN and Beclin 1/GFAP were observed. Fluorescence colors: NeuN: green, GFAP: green and Beclin 1: red (n = 6). Scale bars: 100 μm. RAPA: Rapamycin; SCI: spinal cord injury, GFAP: glial fibrillary acidic protein.

Figure 9| NeuN expression in the dorsal column at 24 hours after SCI. (A) Representative western blots showing levels of NeuN. (B) Relative band densities of NeuN. The densities of the protein bands were analyzed and normalized to β-actin. Data are expressed as the mean ± SD (n = 6; one-way analysis of variance followed by Dunnett’s t-test). *P < 0.05, vs. sham group; #P < 0.05, vs. SCI + vehicle group; ?P < 0.05, vs. SCI + RAPA group. RAPA: Rapamycin; 3-MA: 3-methyladenine; IV: Akt inhibitor IV; SCI: spinal cord injury.

Figure 5 EPO inhibits neuronal ferroptosis via upregulating expressions of xCT and Gpx4. (A) Double-labeled immunofluorescent staining of 4-Hne (red, TRITC)/NeuN (neuronal marker, green, FITC) from injured tissue and relative fluorescence intensity analysis at 14 dpi. (B) Double-labeled immunofluorescent staining of Gpx4 (red, TRITC)/NeuN (green, FITC) from injured tissue and relative fluorescence intensity analysis at 14 dpi. (C) Double-labeled immunofluorescent staining of xCT (red, TRITC)/NeuN (green, FITC) from injured tissue and relative fluorescence intensity analysis at 14 dpi. Nuclei were stained by DAPI (blue). Scale bars: 100 μm. All data are expressed as the mean ± SD (n = 5 in each group). *P < 0.05, **P < 0.01, ***P < 0.001 (one-way analysis of variance followed by Dunnett’s multiple comparisons test). 4-Hne: 4-Hydroxynonenal; Dapi: 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride; dpi: day(s) post injury; EPO: erythropoietin; Fer-1: ferrostatin-1; FITC: fluorescein isothiocyanate; Gpx4: glutathione peroxidase 4; IF: immunofluorescent; NeuN: neuronal nuclei antigen; RSL3: (1S,3R)-RSL3; SCI: spinal cord injury; TRITC: tetramethylrhodamine isothiocyanate; xCT: the solute carrier family 7 member 11.

Figure 10.| Sal promotes increases in dendritic spine density and synaptic-associated protein expression via upregulating the FGF2-mediated cAMP/PKA/CREB signaling pathway.(A) Examples of dendritic spines (the scale bar is 10 μm). (B, C) Density of dendritic spines. (D) Double immunofluorescence staining of PSD95-positive (green) and NeuN-positive (red) neurons from sections in each group on day 7 after MCAO/R (the scale bars are 20 and 10 μm).

Figure 4 Melatonin enhances cell viability in the spinal cord. (A and B) Immunofluorescence staining was performed to visualize and quantify the number of NeuN-positive cells in the anterior horn of the spinal cord at the lesion epicenter. The scale bar represents 200 µm. (C-E) mRNA levels of Bcl-2 and Bax, as well as the ratio of Bcl2/Bax, were evaluated in spinal cord tissue at 7 days post-injury. *P≤0.05. Mel, Melatonin; Veh, Vehicle; 4PP, 4-phenyl-2-propionamidotetralin.

Figure 7 Neurological effects of Mito/M1-BV2 and Mito/M0-BV2 on tMCAO rats. (a) After Mito/M1-BV2 was labeled red and injected into the ischemic cortex of rats, the immunofluorescent staining revealed colocalization of Mito Tracker Red-labeled microglial mitochondria with neurons labeled with anti-NeuN (green). Scale bar: 20 μm. (b) Cerebral sections (n = 5) were stained using 2,3,5-triphenyltetrachloroammonium (TTC). After ischemia for 2 h and reperfusion for 6 h, the infarct volume was calculated. In addition, Ludmila belayev (n = 5) and Zea-longa neurological scores (n = 5) were determined. The data indicate mean ± SD, ∗P < 0.05, and ∗∗∗P < 0.001. (c) The immunofluorescent staining of NeuN-positive cells (green) in ischemic region of tMCAO rats treated with Mito/M0-BV2 and Mito/M1-BV2. NeuN antibody was utilized for neuron staining in ischemic cerebral tissues of tMCAO rats (n = 3). DAPI (blue) served as the nuclear marker. NeuN-positive neuronal counts were quantified. The relative neuron number was determined by calculating the ratio of the number of NeuN-positive cells in groups to the number of NeuN-positive cells in the sham group. Scale bar: 100 μm. Data presented are mean ± SD. ∗∗∗P < 0.001. (d) TEM images of mitochondria in ischemic cerebral tissues of tMCAO rats. Scale bar: 0.5 μm.