(a) Dose-response and (b) time-response effect of CoCl2 on HIF-1α levels in +SA mammary cancer cells grown in culture. +SA cells were seeded at concentration of in 100 mm culture dishes and allowed to attach overnight. The following day, cells were divided into treatment groups and exposed to various concentrations of CoCl2 for 0–24 hr incubation period. Afterwards, cells were isolated with trypsin, and whole cell lysates were prepared for Western blot analysis. Scanning densitometric analysis was performed on all blots done in triplicate and the integrated optical density of each band was normalized with corresponding α-tubulin, as shown in the bar graphs below their respective Western blot image. Vertical bars indicate the normalized integrated optical density of bands visualized in each lane SEM. as compared to the vehicle-treated controls.
