Figure 6

Ras-transformed cells demonstrate increased dependence on mitochondrial respiration. a. Oxygen consumption was measured using 5 × 10⁶ primary (NHBE), immortalized (hT/LT) and Ras-transformed (hT/LT/Ras) bronchial epithelial cells in normoxia (21% oxygen) with a Clark type polarographic electrode. b. 5 × 10⁵ primary bronchial epithelial cells (NHBE), immortalized (hT/LT) and transformed (hT/LT/Ras) cells were cultured in normoxia or anoxia +/- 10 μM rotenone for 24 h. The cells were washed X1, lysed and ATP was measured using a bioluminescent assay and values expressed as % of control. Cells in normoxia without rotenone were used as the control (ATP amounts of controls: NHBE 0.707 ± 0.018, hT/LT 0.734 ± 0.07, hT/LT/Ras 0.7 ± 0.038 pmol per μg protein). c. 5 × 10⁵ NHBE, hT/LT and hT/LT/Ras cells were cultured in normoxia or anoxia with or without rotenone for 24 hrs. Viable and non-viable cells were enumerated by Trypan blue exclusion using a hemacytometer.