Assessment of Microvessel Isolation Protocol by real-time Quantitative Reverse Transcription Polymerase Chain Reaction to Gauge the Level of Capillary Enrichment

Relative quantification of mRNA transcripts in samples generated from whole murine brain (yellow), vessels collected after enrichment (blue), and microvessels collected after enrichment and filtration (pink). mRNA samples were probed for the relative levels of Acta2, Myh11, Cnn1, Ephb4, Nrf2f, Cldn5, and Pecam1, with TATA-binding protein (Tbp) referenced as the loading control. This analysis provided greater insight into the output from each collection technique with respect to microvessel content.

本研究对三类样本的mRNA转录本开展相对定量分析：分别为全小鼠脑组织（标记为黄色）、富集处理后收集的血管（标记为蓝色），以及经富集与过滤两步处理后收集的微血管（标记为粉色）。针对上述mRNA样本，本研究检测了Acta2、Myh11、Cnn1、Ephb4、Nrf2f、Cldn5及Pecam1的相对表达水平，并以TATA结合蛋白（TATA-binding protein, Tbp）作为上样内参。该分析可更深入地解析不同样本收集策略在微血管组分富集效果上的差异。