(A-F) Western blot analysis of protein PAR in EC lysates. (A, B) HUVECs were pre-treated with or without AICAR for 4 hr before the addition of glucose (30 mM) (A) or Ang II (100 nM) (B) and incubated for another 24 hr. (C) HUVECs were treated with or without metformin or glipizide for 6 hr, then incubated with or without 30 mM glucose for 24 hr. (D) HUVECs were treated with or without telmisartan or metoprolol for 6 hr, then incubated with or without 100 nM Ang II for 24 hr. (E, F) BAECs were transfected with flag-tagged wild-type (WT), S177A, or S177D PARP1 plasmids for 24 hr, then incubated with 30 mM glucose (E) or 100 nM Ang II (F) for 6 hr. (G) BAECs were transfected with S177A or S177D PARP1 plasmid. RT-PCR analysis of mRNA level of eNOS, SIRT1, KLF4, MCP-1, and VCAM-1. (H, I) BAECs were transfected with S177A or S177D PARP1 or WT PARP1 treated with or without PJ-34 (3 μM for 6 hr). (H) Western blot analysis and quantification of protein levels. (I) Measurement of PARP1 activity in nuclear extracts of BAECs and NO level in the cultured medium. Data are mean±SD from at least 3 experiments. *p<0.05 compared with controls.