NF-κB/p65 levels are increased and HUWE1 levels are reduced in lungs of SS mice in vivo and in HEM-treated HPAECs in vitro. (A) Whole lung homogenates were collected from littermate control (AA) and SS mice. Lung NF-κB/p65 mRNA or protein levels were measured with real-time qPCR or western blotting and expressed relative to lung mRNA (9S mRNA) or protein (GAPDH). *P < .05 vs AA, n = 4 to 5. (B-C) HPAECs were treated with DMSO vehicle (CON) or HEM (2.5, 5.0, and 10.0 µM) for 72 hours. (D-E) HPAECs were treated with DMSO vehicle (CON) or HEM (5 µM) for 24, 48, and 72 hours. Mean HPAEC NF-κB/p65 mRNA or protein (B) and ET-1 mRNA (C) levels were measured with real-time qPCR or western blotting. Each bar represents the mean NF-κB/p65 or ET-1 level ± SEM relative to GAPDH expressed as fold change vs CON. *P < .05 vs CON, n = 6. (D-E) HPAECs were treated with DMSO vehicle (CON) or HEM (5 µM) for 24, 48, and 72 hours. Mean HPAEC p65 mRNA and protein (D) and ET-1 (E) protein levels were measured with real-time qPCR or western blotting. Each bar represents the mean p65 or ET-1 level ± SEM relative to GAPDH expressed as fold change vs CON. *P < .05 vs CON, n = 6.