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Fig. 9 | Journal of Neuroinflammation

Fig. 9

From: Alpha-synuclein oligomer-selective antibodies reduce intracellular accumulation and mitochondrial impairment in alpha-synuclein exposed astrocytes

Fig. 9

Antibody treatment results in lower extracellular levels of α-syn. Extracellular levels of total α-syn were measured with a sandwich ELISA and immunoprecipitation followed by Western blot. a Co-cultures were exposed to α-syn oligomers or to oligomers pre-incubated with mAb47, mAb49G, or 38E2. Conditioned media analyzed with sandwich ELISA demonstrated that the α-syn levels were dramatically decreased when any of the antibodies were present. The largest reduction in extracellular α-syn levels was seen with mAb47. Data are presented as mean ± SD, and the levels of significance were set to *P < 0.05, **P < 0.01, and ***P < 0.001 (n = 3). b To detect the fraction of α-syn oligomers bound to mAb47, immunoprecipitation was performed on conditioned media from the mAb47-treated sample (24 h). As a positive control, media containing mAb47:oligomer complex that had not been in contact with cells was used (0 h). Western blot on eluted immune complexes displayed a smear of high molecular weight α-syn species, dimer (35 kDa), and monomer bands (14 kDa) in the 0 h control. The 24 h samples showed reduced α-syn signals, as compared to the 0 h control. In addition, the heavy (50 kDa) and light (25 kDa) IgG chains from mAb47 were detected. c Quantification of α-syn bands revealed a clear decrease after 24 h, as compared to 0 h. d The heavy and light chain bands of mAb47 were quantified and displayed only a modest reduction after 24 h (n = 3)

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