(A) Cellular levels of ROS were measured using DCFH-DA in patient-derived or rat-derived cells grown in serum-free conditions. (B) Rat-derived cells and (C) LAM patient-derived cells were treated with 10 nM E2, 10 nM Pg, 10 nM E2 + 10 nM Pg, or vehicle control for 24 hr. Cells were incubated with hydrogen peroxide (0.5 µM) for 30 min prior to the measurement. Cellular levels of ROS were quantified using DCFH-DA. Data was normalized to total cell number. Results are representative of eight independent samples per group from three experiments. (D) Cellular levels of NADPH were measured in LAM patient-derived cells treated with 10 nM E2, 10 nM Pg, 10 nM E2 +10 nM Pg, or vehicle control for 24 hr. Data was normalized to total protein level. (E) Cellular levels of GSH were measured in patient-derived cells treated with 10 nM E2, 10 nM Pg, 10 nM E2 +10 nM Pg, or vehicle control for 24 hr. Data was normalized to total protein levels. Results are representative of three sets of independent samples per group from three experiments. * p < 0.05, ** p < 0.01, Student t test.