Effect of SK inhibition on Nrf2 in human airway epithelial cells (BEAS2B).

<p><b>A.</b> Nuclear extracts from cells treated with increasing concentrations of SK inhibitor SKI-II (0.03 to 3 µM) for 2 h were analysed by immunoblotting for Nrf2 expression and normalized using TBP (fold change vs. NT). *** p<0.0001. <b>B.</b> Expression of antioxidant genes NQO1 and GCLM was determined 24 h after and HO-1 after 8 h treatment with SKI-II. GNB2L1 was used as housekeeping gene. ** p<0.001, * p<0.05. <b>C.</b> Nuclear fractions from cells treated with SKI-II (1 µM) at increasing times (10–120 min) were analysed for Nrf2 expression and normalized using TBP (nuclear). *** p<0.0001,* p<0.05. <b>D.</b> Nuclear fractions from cells treated with SKI-II (1 µM) at increasing times (10–120 min) were analysed for Anti-oxidant Response Element (ARE) binding. *** p<0.0001, ** p<0.001. <b>E.</b> BEAS2B cells were treated with cycloheximide (CXM) and SKI-II (1 µM) at different time points (5 to 60 min) and nuclear extracts were analysed for Nrf2 expression normalized against TBP. Data are representative of 3 independent experiments and are means and S.E. of triplicates.