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. Author manuscript; available in PMC: 2014 Aug 16.
Published in final edited form as: Circ Res. 2013 Jul 12;113(5):10.1161/CIRCRESAHA.113.301792. doi: 10.1161/CIRCRESAHA.113.301792

Figure 7. Depletion of MGP allows for multipotency and osteoinduction in human aortic endothelial cells (HAECs).

Figure 7

(A) Expression of multipotent markers Sox2, Nanog and Oct3/4 in HAECs after transfection of scrambled control siRNA (SCR) or MGP siRNA only (lane 1, 2), siRNA transfection with BMP4 treatment (lane 3, 4), siRNA transfection with Noggin treatment (lane 5, 6), or siRNA transfection with BMP4 and Noggin treatment (lane 7, 8), as determined by immunoblotting. (B) Co-expression of CD31 and SSEA-3, or CD31 and SSEA-4 after transfection of scrambled control siRNA or MGP siRNA without additional treatment (left), and with BMP4 treatment (right), as determined by FACS analysis. (C) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, Osterix (OSX), early SMC markers SM22α, α-SM actin (αSMA), and late SMC marker SM-myosin heavy chain (SM-MHC) (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), osteogenic media (O) or both (B+O), as determined by immunoblotting. (D) Staining for alkaline phosphatase (ALP), and mineral (Alizarin Red and Von Kossa staining) in HAECS treated as described in (c). (E) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, OSX, SM22α, αSMA, and SM-MHC (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), high glucose medium (G) or both (B+G), as determined by immunoblotting. (F) Staining for ALP and mineral in HAECS treated as described in (E).