BMECs were plated on 24-well plates in EC medium or EC medium conditioned by primary brain pericytes or hPSC-derived pericyte-like cells. After 48 hours, BMECs were fixed and stained for occludin as described above. Images were acquired from five wells per experimental condition. To quantify tight junction continuity, images were blinded and the area fraction index was determined using FIJI as previously described (78). In addition, images were blinded, and the number of frayed junctions (Fig. 4D) was manually counted.

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