HIF-1α protein expression and measurement of oxygen consumption in CHOK1 cell line exposed to different doses of rotenone.
(A) Representative immunoblot of HIF-1α showing dose response characteristic for the effect of rotenone on induction by hypoxia. Levels of HIF-1α in extracts of CHO-K1 cells after 4 hour exposure to normoxia (21% O2, N) or hypoxia (0.1% O2, H) at different doses of rotenone (0 to 10 μM). (B) Data are HIF-1α signals (mean ± 1 SD; n = 3) normalized to the HIF-1α signal from control cells in hypoxia. (C,D) Polarographic measurements of oxygen consumption in CHO-K1 cells exposed to rotenone. (C) Dose response for rotenone (0, 0.1 μM, 1 μM). Cells treated for 4 hours in normoxia (21% O2). (D) Time course. Cells treated with 0.1 μM rotenone for 30 minutes or 4 hours in normoxia (21% O2). Measurements were performed in triplicate. Data are mean ± 1 SD.
