BMVEC recovered from BBB model have extensive junctional proteins. BMVEC were collected from the BBB model using Try-pLE and junctional proteins examined by flow cytometry. (A) FSC-H and FSC-A were used for the identification of singlets (indicated within the gated population) and doublet discrimination. (B) The single cells were then examined by SSC-A and FSC-A to develop the BMVEC gate. FITC or PE fluorescent signal was used to examine (C) PECAM-1, (D) ALCAM, (E) CD99, (D) PrPC and the respective isotype matched negative controls in one representative experiment. The MFI is denoted. (G) The MFI of PECAM-1, ALCAM, CD99, and PrPC (open bars) or their respective isotype-matched negative controls (closed bars) were analyzed in six independent experiments. Data are represented as mean ±standard deviation. Significance was determined by a two-tailed paired t-test. *P <0.05. **P <0.01. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]