Extended Data Fig. 2. Cholesterol binding stabilizes VCAM-1.

(a) Western blot for VCAM-1 in HUVECs cultured in media containing either 10% FBS with simvastatin or 1% FBS with simvastatin for 24 h before stimulation with LPS (100 ng/ml) for 0–24 h. (b) VCAM-1 western blots in cells stably overexpressing human VCAM-1 and cultured in media containing either 10% FBS or 1% LPDS with simvastatin for 16 h. (c) VCAM-1 western blots in the NP-40 resistant fraction of HUVECs stably overexpressing FLAG-VCAM-1 and cultured in media containing 1% LPDS with simvastatin for the indicated times. In the last two lanes, 100 μM MβCD-cholesterol or LDL (50 ug/ml) was added for the last 2 or 4 h of the 24 h LPDS chase, respectively. (d) Total plasma cholesterol in male WT mice 6 h after receiving i.v. infusions of either saline or LDL (n = 7 saline and 8 LDL). Data are represented as mean ± SEM with individual mice noted as dots.