A) Menadione (12.5μM) increases intracellular levels of 2-HE in LPS-treated FPAEC incubated in hypoxia: 2-HE levels were quantified in FPAEC by HPLC after n-Butanol extraction. 2-HE levels increased by greater than two-fold when LPS treated FPAEC were pre-treated with menadione (0.09 vs. 0.25 nmol 2-HE/mg protein). Hypox + LPS; LPS treated FPAEC incubated in hypoxia, Hypox + LPS + Menadione; LPS and menadione treated FPAEC incubated in hypoxia. * - P<0.001.
B) Pre-treatment of FPAEC incubated in hypoxia with menadione abolishes differences in LPS-induced apoptosis between normoxia and hypoxia: Quantification of apoptosis in FPAEC treated with LPS (10μg/mL) and incubated under normoxic or hypoxic conditions (5% O2) for 48 hours with or without 12.5μM of menadione. Cells detected in early and late apoptosis for each condition were pooled and averaged from six independent experiments. Menadione alone; FPAEC treated with menadione alone, Norm + LPS; LPS treated FPAEC incubated in normoxia, Hypox + LPS; LPS treated FPAEC incubated in hypoxia, Hypox + LPS + Menadione; LPS and menadione treated FPAEC incubated in hypoxia. § - P<0.001, * - P=0.003, ** - P=0.39.