Investigating NF-κB Signaling in Smooth Muscle Cells

Human pulmonary artery (hPASMC) (Lonza CC-2581) from three commercial donors and four primary donors, including those with and without disease-related BMPR2 mutations, and human aortic smooth muscle cells (hAoSMC) (Lonza CC-2571) from two donors were maintained in smooth muscle cell media SmGM-2 including bullet kit supplements (Lonza CC-3182). All experiments were performed between passages 4 and 8. Quiescent media (Dulbecco’s modified Eagles medium (Lonza 12-604) with 0.2% (v/v) fetal bovine serum ((FBS) Lonza 14-401) was added for 48 h prior to transfection with siRNA/reporter plasmid (Qiagen Cignal NFkB luciferase reporter) and/or 6-h stimulation with IL-1ß at 10 ng/mL or BMP4 at 20 ng/mL.^{13 (link),14 (link)}