Synaptophysin Antibody - #AF0257

Fig. 3 OTUD1 knockout alleviates cognitive dysfunction in SAE mice by reducing neuronal and synaptic damage. (A) Experimental design for animals in vivo. (B) Violin plots displaying the expression of Otud1 in microglial subclusters. (C) OTUD1 expression in hippocampus was analyzed by western blot (n = 3/group). (D) The preference index of the four groups of mice in the NORT (n = 8/group). (E) Escape latency, (F)time spent in the target quadrant, and (G) numbers of crossings through the platform were recorded in each group (n = 8/group). (H) Trajectory plot of spatial exploration in MWMT. (I-K) Neuronal damage of the hippocampal region was assessed by HE staining and Nissl staining (n = 5/group). Scale bar, 50 μm. (L) Synaptic structural changes were observed through TEM. SV, Synaptic vesicles; SC, Synaptic cleft; PSD, Postsynaptic density. Scale bar, 100 nm. (M-N) Image analysis of the thickness of PSD and the width of the synaptic cleft (n = 3/group). (O-P) The levels of PSD95 and SYP in hippocampus were measured by western blot (n = 3/group). *p 

Fig. 2 Erbin alleviates SAE by attenuating neuronal damage, improving the synaptic structure, and increasing synaptic protein expression. A–C Neuronal damage of the hippocampal region was assessed by Nissl staining. Scale bar, 50 μm. D, E Image analysis of the thickness of PSD and the width of the synaptic cleft. F Synaptic structural changes were observed in TEM. Scale bar, 50 nm. SV, Synaptic vesicles; SC, Synaptic cleft; PSD, Postsynaptic density. G–J Levels of PSD95, SYP, and SYN1 in hippocampal tissue were measured by western blot. Data are representative of three independent experiments. *p 

Fig. 3. The cerebral protection effects of FMT in mice against age-associated proteins expression levels. (A, B) Quantified protein levels of the cell cycle arrest related proteins of p53, p21, p16/p14, and Rb in mice hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA followed by the Dunnett T3 test for comparison of multiple groups, *P < 0.05, **P < 0.01, and ***P < 0.001. (C, D) Quantified protein levels of the DNA damage-related protein ATM, and the cognitive-related proteins synapsin I, synaptophysin and PSD95 in mice hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA followed by the Dunnett T3 test for comparison of multiple groups, *P < 0.05, **P < 0.01, and ***P < 0.001. (E, F) Quantified protein levels of the cell senescence-related proteins CREB, p-CREB, ERK, p-ERK, AKT, p-AKT in mice hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA followed by the Dunnett T3 test for comparison of multiple groups, *P < 0.05, **P < 0.01, and ***P < 0.001. (G, H) Quantified protein levels of the c-H2AX and TP53BP1 proteins in bone marrow mesenchymal stem cells. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA followed by the Dunnett T3 test for comparison of multiple groups, *P < 0.05, **P < 0.01, and ***P < 0.001. (I, J) Quantified protein levels of the β-galactosidase, c-H2AX, and TP53BP1 proteins in in mice hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA followed by the Dunnett T3 test for comparison of multiple groups, *P < 0.05, **P < 0.01, and ***P < 0.001.

Figure 3. The effect of Sig-1R agonist on synapse change in the hippocampus of mice with T1DM. (a–c) Representative Western blots and quantitation of PSD-95 and Syp protein expression in CON, CON + PRE-084, STZ, and STZ + PRE-084 mice (n = 6). (d–g) Representative images of synaptic ultrastructure (d), quantitative analysis of PSD length (e) and width (f), and synaptic cleft (g) (n = 85–90; 3 animals per group). The scale bars are 500 and 200 nm. (h) Golgi staining images of the neuronal morphology. The scale bar is 50 μm. (i) Representative three-dimensional reconstruction images of different types of the spine in hippocampal neurons, including stubby (red), mushroom (green), thin (blue), and filopodia (purple) spines. The scale bar is 2 μm. (j–n) Quantitation of density of different spines, including total (j), stubby (k), mushroom (l), thin (m), and filopodia (n) spines (n = 20; 3 mice per group). The data were presented as mean ± SEM and analyzed via one-way ANOVA with Tukey’s multiple comparison tests, Welch’s ANOVA test followed by Dunnett’s T3 multiple comparison tests, and Kruskal-Wallis test followed by Dunn’s multiple comparison tests. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Sig-1R, sigma-1 receptor; T1DM, type 1 diabetes mellitus; PSD, postsynaptic density; Syp, synaptophysin; CON, control mice; STZ, mice with T1DM.

Figure 3. Protective effects of ginseng and FMT in mice against age-associated protein expression levels. (A,B) Quantified protein levels of cell cycle arrest-related proteins of p53, p21, p16/p14, and Rb in mouse hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA, followed by the LSD post hoc test or Dunnett’s T3 test for the comparison of multiple groups; *p < 0.05, **p < 0.01, and ***p < 0.001. (C,D) Quantified protein levels of the DNA damage-related protein ATM and the cognitive-related proteins synapsin I, synaptophysin, and PSD95 in mouse hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA, followed by the LSD post hoc test or Dunnett’s T3 test for the comparison of multiple groups; *p < 0.05, **p < 0.01, and ***p < 0.001. (E,F) Quantified protein levels of the cell senescence-related proteins CREB, p-CREB, ERK, p-ERK, AKT, and p-AKT in mouse hippocampus tissues. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA, followed by the LSD post hoc test or Dunnett’s T3 test for the comparison of multiple groups; *p < 0.05, **p < 0.01, and ***p < 0.001. (G,H) Quantified levels of the c-H2AX and TP53BP1 proteins in bone marrow mesenchymal stem cells. Data are shown as the mean ± SD, n = 3 per group. Significance was determined by one-way ANOVA, followed by the LSD post hoc test or Dunnett’s T3 test for the comparison of multiple groups; *p < 0.05, **p < 0.01, and ***p < 0.001.

Figure 5 Western blot analysis. The expressions of synaptic plasticity proteins were determined by western blot (A), including PSD-95 (B), GAP-43 (C), and SYN (D). The values represent the mean ± SEM, n = 5. *p < 0.05, **p < 0.01 vs. CON, #p < 0.05, ##p < 0.01 vs. MS. CON, control; MS, maternal separation; CUMS, chronic unpredictable mild stress.