Figure 3.

Lectin staining of human brain microvessel endothelial cells (hBMEC) under normoxic and OGD conditions. Ai. Effect of IGF-1 and its receptor antagonist on intercellular spaces: Photomicrographs of lectin-stained cultures show that the even, confluent monolayer seen in normoxia is changed under OGD to smaller, densely-stained cells with large intercellular spaces. Intercellular spaces were completely abrogated in IGF-1 treated cultures and this was reversed with concurrent exposure to JB-1. Aii Histogram shows average intercellular space estimates using the Canny Edge algorithm. B ii. Effect of IGF-1 and signaling pathway inhibitors on intercellular spaces: Photomicrographs of lectin-stained cultures confirm that IGF-1 abrogates intercellular spaces caused by OGD. All experimental groups are compared to the normoxia group. Concurrent treatment with IGF-1+LY294002 treated cultures shows greater gaps between cells compared to IGF-1 group, while concurrent treatment with Rapamycin is no different from IGF-1 alone. Each data point is the average of 3 technical replicates, and shown here is the average from 3 separate experiments. *p<0.05. Bar: 100 um