Autoantibodies Targeting AT1 Receptor from Patients with Acute Coronary Syndrome Upregulate Proinflammatory Cytokines Expression in Endothelial Cells Involving NF-κB Pathway

Figure 6

Activation of NF-κB by EMSA analysis after AT1-AAs stimulation in endothelial cells. After incubating cells with angiotensin II (0.1 μM) for 30 min and AT1-AAs (0.1 μM) for 0, 15, 30, and 60 min, activity of NF-κB in endothelial cells was determined by EMSA analysis (panel (a)). Lane 1: probe alone without nuclear extract; lane 2: incubation with angiotensin II for 30 min; lane 3: control (before incubation with AT1-AAs); lane 4: incubation with AT1-AAs for 15 min; lane 5: incubation with AT1-AAs for 30 min; lane 6: incubation with AT1-AAs for 60 min; lane 7: mutant oligonucleotide to test the specificity of the method; lane 8: 200-fold molar excess of unlabeled NF-κB oligonucleotide as a competitive inhibitor. Data were summarized from 5 independent experiments (panel (b)); versus control group (lane 3).

(a)

(b)