To detect vascular Notch3 extracellular domain aggregates in CADASIL, we developed a novel dot-blot assay with both autopsy and biopsy skin samples. We obtained samples from 11 patients with CADASIL and 12 control patients, and we performed dot-blot analyses by using sequential biochemical tissue extractions with three different antibodies against specific regions of the Notch3 extracellular domain. We also analyzed clinical features and vascular accumulations of Notch3 by immunohistochemistry. Via the dot-blot assay with the antibody against the C-terminal region of the Notch3 extracellular domain, we successfully detected Notch3 extracellular domain aggregates in skin tissue homogenates obtained from patients with CADASIL. Our novel method may therefore aid the diagnosis of CADASIL.

中文翻译：

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用于检测 CADASIL 患者血管 Notch3 聚集体的新型斑点印迹法

为了检测 CADASIL 中的血管 Notch3 细胞外结构域聚集体，我们开发了一种新的斑点印迹法，用于尸检和活检皮肤样本。我们从 11 名 CADASIL 患者和 12 名对照患者中获得了样本，并通过使用针对 Notch3 细胞外结构域特定区域的三种不同抗体的连续生化组织提取物进行了斑点印迹分析。我们还通过免疫组织化学分析了 Notch3 的临床特征和血管积累。通过使用针对 Notch3 细胞外结构域 C 末端区域的抗体进行斑点印迹分析，我们成功地检测到了从 CADASIL 患者获得的皮肤组织匀浆中的 Notch3 细胞外结构域聚集体。因此，我们的新方法可能有助于 CADASIL 的诊断。