Western Blot Analysis of Protein Targets

Western blot was performed as described [41 (link)]. Antibodies against p53, p21, Egr-1, IRP2 (The anti-IRP2 antibody used in Figs. 3–5 and Fig. 6 have different lot number), FTH1, TfR1, and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-mouse IRP2 antibody was a generous gift of Elizabeth Leibold (University of Utah) [53 (link)]. Anti-mouse p53 (1C12) and anti-PARP antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against FDXR were purchased from Abcam (Cambridge, MA, USA). HRP-conjugated secondary antibodies against rabbit and mouse IgG were purchased from BioRad (Hercules, CA). The immunoreactive bands were visualized by enhanced chemiluminescence (Thermo Fisher Scientific Inc, Carlsbad, CA) and quantified by densitometry with the BioSpectrum® 810 Imaging System (UVP LLC, Upland, CA).