Effects of 150 µM CoCl2 (hypoxic, but not cytotoxic dose) alone and in combination with compound 44 (44), on MAPK cascade signaling proteins in +SA mammary tumor cells. +SA cells were seeded at concentration of in 100 mm culture dishes and allowed to attach overnight. The following day, cells were divided into groups and exposed to their respective treatments for a 24 hr incubation period. Afterwards, whole cell lysates were prepared for Western blot analysis or MEK, phospho-MEK 1/2 (p-MEK 1/2, Ser221/217), ERK1, ERK2, and phospho-ERK 1/2 (p-ERK 1/2, Thr202/Tyr204). 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 hypoxic CoCl2-treated control group.
