Figure 2. Anthracyclines interfere with the transcription of HIF-1α and HIF-2α.

A.-B. Hep3B cells were exposed to CoCl₂ in the presence of DMSO (Control) or 0.625 μM EPI, DOX, or IDA for 16 hours. HIF-1α and HIF-2α mRNA levels were measured by RT-qPCR. Anthracycline treatment under CoCl₂ was compared with CoCl₂ treatment alone. *P < 0.05. C. HIF-1α and HIF-2α protein levels, in cells treated with CoCl₂ in the presence of DMSO or EPI, DOX, or IDA, were determined by immunoblot assays. β-Actin was used as the internal control. D. Knock-down effects of HIF-1α and HIF-2α siRNA were verified by western blot. β-Actin was performed as the internal control. E. siHIF-1α and siHIF-2α were used to knock down gene expression and their inhibitory effects on cell migration were observed. Bar = 100 μm. F. Cells were incubated with or without CHX (100 μM) and IDA (0.625 μM) for the indicated incubation times. Protein level changes of HIF-1α and HIF-2α were measured by western blot. G. Quantification and half-life of HIF-1α and HIF-2α under IDA treatment with DMSO was used as the control vehicle.