Extended Data Fig. 3. PM cholesterol accessibility increases during EC activation and hypercholesterolemia.

(a) HUVECs stably expressing FLAG-VCAM-1 and cultured in media containing 1% LPDS with simvastatin overnight were treated with or without neutral sphingomyelinase for 2 h before immunoblotting for VCAM-1. (b) En face imaging of aortas from female mice that had been perfused with cholesterol-binding mutant versions ALOD4–647. Samples were co-stained with VCAM-1 (red), Ve-cadherin (green), and DAPI (blue). Scale bar, 30 μm. (c) ALOD4–488 binding to en face aortas of either male WT mice fed a chow diet or LDLR knockout mice that had been fed a Western diet for 20 weeks to induce atherosclerosis. Samples were co-stained with DAPI (blue). Scale bar, 30 μm.