SNAP25 Antibody - #AF0254

Figure 7 Ser727 of STAT3 increased its interaction with PKCε. (a) Endogenous PKCε was immunoprecipitated from cell lysates, and immune complexes and total cell lysates were analyzed by Western blot analysis with PKCε and STAT3 antibodies. Endogenous immune PKCε/STAT3 complexes were detected in the rat spinal cord tissues. (b) IL-6 promoter-firefly luciferase reporter plasmid (0.5 μg), PKCε (1 μg), and STAT3 (1 μg) were cotransfected overnight into HEK293 cells. The transfected cells were incubated with 1 μg/mL lipopolysaccharide (LPS) for 12 h. Interleukin-6 promoter activity increased by STAT3 was further enhanced by PKCε and STAT3, indicating that PKCε improved the ability of STAT3 to bind to IL-6 promoter. Data are shown as means ± SD (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; one-way ANOVA followed by Bonferroni tests. (c, d) HKE293 cells were transfected with GFP, GFP-PKCε, Flag, Flag-STAT3, and phosphomimetic and dephosphomimetic mutants of STAT3, and then, immunoprecipitants were assayed. Protein complexes were detected using an anti-GFP antibody (c), and then, relative PKCε binding to STAT3 was quantified (d). Data are presented as means ± SD (n = 3). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; one-way ANOVA followed by Bonferroni tests.

FIGURE 5 Ubiquitin and synaptic protein expression changed in isoflurane-exposed aged mice. (A) Ubiquitin expression was decreased in the cortex brain tissue in the ISO-15 group, while there was no significant difference in synaptic protein SNAP25 expression in the cortex (n = 6, *p < 0.05). (B) In the brain tissue of cortex, there was no significant difference in the expression level of synapsin I and the phosphorylation level of synapsin I S9 and S605 between CON group, ISO-12.5 group, and ISO-15 group. In the brain tissue of the hippocampus, the expression level of synapsin I and the phosphorylation level of synapsin I S9 and S605 were decreased in both the ISO-12.5 group and ISO-15 group, compared with the CON group, and the expression level of CHIP had a dramatic decline in the ISO-15 group (n = 6, *p < 0.05).

FIGURE 6 | Decreased CHIP expression altered synaptic protein expression and phosphorylation in N2a cells. (B) Statistical bar chart to show that CHIP, synapsin I, SNAP25, and PSD95 expression were present (n = 6, **p < 0.01, ***p < 0.001).