The increase in Ndrg1 and TfR1 mRNA following Fe deprivation with DFO and 311 can be reversed with the Fe donor, ferric ammonium citrate. MCF-7 cells were incubated with control medium (minimum essential medium; MEM), DFO (250 μM), or 311 (25 μM) for 14 hours at 37°C (primary incubation). This medium was then removed, and the cells were then reincubated for 14 hours at 37°C with either control media (MEM) or media containing either the Fe donor ferric ammonium citrate (FAC; 100 μg/mL), DFO (250 μM), or 311 (25 μM) (secondary incubation). The expression of (A) Ndrg1 and (B) TfR1 mRNA levels (positive control) were then assessed by using semiquantitative RT-PCR (described in “Materials and methods”). Densitometry was performed, and gene expression was then calculated relative to the β-actin control. Results show a typical experiment from 3 performed.