(A)
BAD expression in pulmonary arterial endothelial cells (PAECs) was quantified after AREG silencing (left panel) and recombinant Amphiregulin treatment (right panel). (B, C, F) PAECs were transfected with either scrambled siRNA (siCTL) or siRNA against BAD (siBAD) and placed in hypoxia for 24 h. (B, C) PAECs apoptosis was assessed by measuring caspase 3+ and caspase 3 MFI cells by flow cytometry (B), and tube formation ability was determined by a Matrigel assay (C). (D, E) PAECs were transfected with either scrambled siRNA (siCTL), siRNA against AREG (siAREG) or siRNA against both AREG and BAD (siBAD/AREG) and placed in normoxic conditions for 24 h. (D, E) Apoptosis (D) and tube formation (E) were examined. (F)
IFNB, IL1B, IL6, and TNFA expression was assessed by qRT-PCR. n = 5 replicates per condition. Data are shown as mean. *P < 0.05, ***P < 0.005, ****P < 0.001.