Figure 1.

Ethanol stimulates Notch signalling in human coronary artery endothelial cells (HCAEC). (a) Notch receptors 1 and 4 (N1, N4), and target gene hrt2 and hrt3 mRNA levels determined by qRT-PCR in HCAEC treated (24 hr) with or without ethanol (EtOH, 25 mM), Delta like ligand 4 (DLL4, 3 μg/ml), or Serum amyloid protein A1 (SAA1, 1 μM). Data are mean ± SEM, n=3–4, #p<0.05, *p<0.01 vs control. (b) Representative Western Blots for N1ICD, N4ICD, hrt2 and hrt3 in HCAEC treated as indicated. β-actin was used as a loading control. (c) Ethanol stimulates γ-secretase activity in HCAEC. Gamma-secretase activity in solubilized HCAEC membranes was measured using a fluorogenic peptide substrate (C-terminal fragment of β-APP). DAPT was used as a control γ-secretase inhibitor (GSI). Data are mean ± SEM, n=4. *P<0.05, **P<0.01 vs C; #P<0.05 +/− DAPT.