Research Paper Volume 14, Issue 3 pp 1292—1306

Figure 8. Kir6.1 upregulation was induced via the ROS/NF-κB-mediated pathway. (A) Flow cytometry analysis of the levels of ROS in CSMCs treated with vehicle (PBS solution) or VF (200 ng/mL), with or without NAC (1 mmol/L) for 24 h. Left panel: Representative image; right panel: Quantitative analysis from three independent experiments. Positive control: Rosup, 50mg/ml. Control: No treatment. (B) Upper panel: Representative double immunofluorescence staining of NF-κB/P65 activity in CSMCs treated with vehicle (PBS solution) or VF (200 ng/L) for 0–60 min. Nuclei were identified by DAPI staining. Lower panel: Western blotting analysis of phosphorylated IκB-αin CSMCs treated with vehicle (PBS solution) or VF (200 ng/L) for 0–60 min. (C) Representative double immunofluorescence staining of NF-κB/P65 activity in CSMCs treated with vehicle (PBS solution) or VF (200 ng/L), with or without NAC (N-acetyl-L-cysteine, 1 mmol/L), for 1 hour. Nuclei were identified by DAPI staining. (D) Western blotting analysis of Kir6.1 subunit protein in CSMCs treated with vehicle (PBS solution) or VF (200 ng/L), with or without NAC (1 mmol/L) or BAY 11-7082 (10 mg/mL), for 1 hour. Upper panel: Representative image; Lower panel: Quantitative analysis from three independent experiments. (E) Measurement of contraction of CSMCs treated with vehicle (PBS solution) or VF (200 ng/L), with or without NAC (1 mmol/L) or BAY 11-7082 (10 mg/mL), for 45 min by scanning micrometry. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001.