Figure 2.

Brusatol decreases HIF-1α protein stability in colorectal cancer cells under hypoxia. (A and B) RKO (A) and HCT116 (B) cells were incubated with or without 100 nM brusatol and exposed to 20% or 0.5% O₂. After an 8 h incubation, cells were harvested at the indicated times. qPCR was used to amplify HIF-1α; 18S rRNA was used as the internal control. (C and D) RKO (C) and HCT116 (D) cells were exposed to 0.5% O₂ for 2 h, incubated with 10 μg/ml CHX for 3 h, and then harvested at the indicated times. HIF-1α protein levels were examined by immunoblot analysis; β-actin was used as an internal control.