FIG. 5.
Activation of NF-κB and expression of ICAM-1 by GPCR-linked mROS. (A) Protein binding to the NF-κB consensus sequence following thrombin stimulation in the presence or absence of BAPTA and MnTBAP. (B) NF-κB transcriptional activity in response to thrombin or TNF-α (10 ng/ml) as assessed by luciferase reporter assay. Luciferase reporter constructs were normalized to β-galactosidase activity. (C) Overexpression of MnSOD abolished thrombin-induced NF-κB activity. (D) Global O2·− production is assessed by HE fluorescence in response to both thrombin (500 mU/ml) and AA (20 μM) following 10 min of stimulation. (E) Quantitation of nuclear HE fluorescence increase in response to AA and thrombin. (F) VCAM-1 and ICAM-1 expression following thrombin or TNF-α stimulation in the presence of MnTBAP (50 μM), BAPTA (25 μM), or the proteosomal inhibitor MG132 in HPMVECs.