Phospho-Synuclein alpha (Ser129) Antibody - #AF3285

Fig. 5 APROM2 reprograms neuronal protein functions via de novo PTMs to fuel synaptic function. a Schematic illustration of the de novo PTM strategy of neuronal proteins using APROM2. APROM2 with protein phosphatase-like characteristics can dephosphorylate phospho-proteins, and thus restore the function of neuronal proteins. Created with BioRender.com. b Top, representative confocal laser scanning microscopy (CLSM) images of p-α-syn in primary neurons after different treatments. Bottom, 3D mapping of intracellular fluorescence. c Mean fluorescence intensity of p-α-syn in primary neurons after different treatments (n = 5 biologically independent cultures). Quantitative analysis (d) and representative CLSM images (e) of synaptic vesicle function indicated by FM1-43 after different treatments (n = 5 biologically independent cultures). Immunofluorescence and quantification analysis of the colocalization of α-syn with VMAT2 (f, h) or VAMP2 (g, i) in primary neurons after different treatments (n = 5 biologically independent cultures). j Microscopy images showing the neurite branches and neuronal connectivity of primary neurons after different treatments. k Intracellular ROS levels in primary neurons after different treatments. l, m Mitochondrial membrane potential (∆ψ) is indicated by JC-1 staining (l) and quantified by normalized JC-1 aggregates/monomers ratio (m) (n = 3 biologically independent cultures). JC-1 monomers (green) represent low ∆ψ, and JC-1 aggregates (red) represent high ∆ψ. n Protective effect of APROM2 and CeNPs on MPP+ treated cells (n = 5 biologically independent cultures). o Schematic illustration of APROM2 that reprograms α-syn function by the de novo PTM strategy and protects mitochondria for fueling synaptic function. APROM2 directly modulates p-α-syn by cleaving the phosphate monoester bond, thus, α-syn regains biological functions of binding to VMAT2 and VAMP2. In addition, APROM2 protects mitochondria against ROS to maintain presynaptic energy homeostasis. All the data are presented as means ± s.e.m. Statistical significance was analyzed by one-way ANOVA with multiple comparisons test. Source data are provided as a Source Data file.

Figure 3 Exogenous GRP78 prevents abnormal accumulation of α-syn phosphorylated at S129 (pS129) in nigral tissue in rat model of Parkinson’s disease. Nigral content of (a) soluble form of α-syn, (b) α-syn phosphorylated at S129 (pS129), (c) phosphorylated to soluble α-syn ratio. Western blot analysis of nigral tissue was conducted with the antibodies against soluble and S129-phosphorylated forms of α-syn. Anti-GAPDH antibody staining was used as the loading control. (d) Representative Western blots are shown in panel. The results are presented as percentages of the control (100%) in (a–c) panels. Bar charts indicate mean values with standard errors. The dots, squares, triangles and rhombus show individual values per rat. Two-way ANOVA test followed by Tukey’s post hoc analysis were performed to determine the effects of GRP78 therapy. Asterisks indicate significant differences between groups according to Tukey’s post hoc tests