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. 2019 Feb 14;12(3):474–487. doi: 10.1016/j.stemcr.2019.01.009

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© 2019 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Immunofluorescent Staining

(A) IMR90-4-derived BMECs labeled for occludin and claudin-5 at day 1 under static conditions. Nuclei are counterstained with Hoechst.

(B) IMR90-4-derived BMECs labeled for F-actin at day 1 under static conditions and day 14 under perfused conditions.

(C and D) HUVECs, μVas, and IMR90-4-derived BMECs labeled for VE-cadherin and claudin-5 at day 7 under static (C) and perfused (D) conditions. Each individual image reflects a summative z projection of individual confocal images without additional processing to flatten images. For each fluorescence channel, the intensity scale is held constant across all samples. As a result, in some images there is a perceived decrease in fluorescence signal for points farthest from the objective that is reflective of channel curvature and slight variations in gelatin topology rather than inherent signal.

Scale bars, 50 μm.