Characterization of iMAEC lines by Dil-Ac-LDL staining. iMAEC lines including wild-type (iMAEC-WT), Caveolin-1 knockout (iMAEC-cav1), eNOS knockout (iMAEC-eNOS), and p47phox knockout (iMAEC-p47), were cultured. (A) Total cell lysates were collected from iMAEC lines or control cells including HUVEC, 3T3, and RASMC. Western blotting was performed using specific antibodies against Flk-1, eNOS and Cav-1. Actin serves as an internal loading control. (B) iMAECs were incubated with Dil-Ac-LDL (10 μg/mL) for 4 h, and images were taken by fluorescence microscopy. HUVEC served as positive control while 3T3 and RASMC served as negative controls. Scale bar: 50 μm. (C) Graph shows the cell shape index of HUVECs, iMAECs and primary MAECs. For cell shape index calculation, 25 cells were chosen randomly from each group and analyzed by ImageJ software. Data represent means ± standard error.