Fig. 7

Requirement of NFκB signaling for TGFβ induced Type II EMT. a Cytoplasmic and nuclear extracts from hSAECs with or without EMT were isolated. 100 μg of cytoplasmic or nuclear extracts were processed for Western blot using anti-RelA Ab (upper panel). Lamin B and tubulin were detected as loading control Abs respectively. Low left panel: epithelial cells or mesenchymal cells were stimulated with 25 μg/ml TNF for 1 h and processed for RelA Western blot. Lower right panel: Quantification of nuclear RelA from Odyssey infrared imager. * P < 0.05 compared with control epithelial cells. b Wild-type (WT) or RelA^−/− mouse embryonic fibroblasts (MEF) were incubated with TGFβ for 0, 1, 2, 3 days respectively. Total RNA was extracted and expression of EMT core transcription factors (SNAIL1 and ZEB1) and NF-κB-dependent genes (IL-6) were measured by Q-RT-PCR. The data shown is from n = 3 independent experiments. c Epithelial cells were stimulated with TGFβ for various times in the absence or presence of a small molecule IκB kinase inhibitor (BMS-345541, 1, 3, and 10 μM respectively). The expression of SNAIL1, Twist1, VIM, ZEB1, FN1 and IL-6 were measured by Q-RT-PCR. The data shown are from n = 3 experiments