Extended Data Fig. 5. Nonvesicular cholesterol transport regulates inflammatory signaling in ECs.

(a) Western blots for VCAM-1 in HAECs treated with (si)Control or (si)Aster-A and cultured in media containing 5% FBS with simvastatin overnight before exposure to LPS for the indicated times. (b) Western blots in HUVECs incubated with or without 100 μM MBCD-cholesterol for 1 h before fractionation of cells into Triton-X100 resistant or Triton-X100 soluble domains. (c) qPCR analysis in cells cultured in LPDS with simvastatin overnight before being loaded with or without the same media containing 20% FBS for 4 h. Target genes were normalized relative to 36B4. (d) qPCR for VCAM1 relative to 36B4 in HAECs cultured in either 10% FBS or 1% LPDS with simvastatin overnight before being stimulated with LPS (100 ng/ml) for 0–8 h. (e) Gramd1a mRNA in isolated hepatic ECs from F/F or ECKO mice. (f) Western blot for Aster-A in isolated hepatic ECs from F/F and ECKO mice. (g) Gramd1a mRNA in liver tissue from F/F or ECKO mice. n = 3 F/F and 3 ECKO. Data are represented as mean ± SEM.