(a) The expression of stem-cell markers was examined by immunoblotting with densitometry in HAECs treated with medium containing elastase 1, 2 (ELAs) and kallikrein 1, 5, 6 (KLKs), and transfected with siRNA (si) to each of the stem-cell markers individually. Ctr: control. SCR: scrambled siRNA. (b) HAECs were depleted of MGP by siRNA (si) and treated with high glucose and BMP-4, then treated with control (lane 1–3), DFP (lane 4–5), serpina1 (lane 6–7), or a combination of DFP, serpina1, and lentivirus (Lv) expressing Sox2 (lane 8–11). Expression of stem-cell markers was examined by immunoblotting with densitometry. (c–f) MGP-depleted HAECs were transfected with siRNAs (si) to elastase (ELA) 1, 2 and kallikrein (KLK) 1, 5, 6, or Sox2, and then treated with control (C), BMP-2 (B), glucose (G), or a combination. Expression of Cbfa1 and Osterix (OSX) was determined by immunoblotting with densitometry (c and d). Calcium accumulation was determined by Alizarin Red and Von Kossa staining (e and f). SCR: Scrambled siRNA.