The Loss of HIF1 Leads to Increased Susceptibility to Cadmium-Chloride-Induced Toxicity in Mouse Embryonic Fibroblasts
Figure 2
Hypoxia signaling and cytotoxicity to CdCl2. (a) Wild-type (WT) cells were left untreated (Ctrl) or exposed to 150 μM CoCl2 (Co2+), 1, 5 or 10 μM CdCl2 (Cd2+) or 150 μM CoCl2 and 10 μM CdCl2 for 24 hours. HIF1α −/− cell extract treated with 150 μM CoCl2 was used as a negative control. Nuclear protein was extracted and separated by SDS-PAGE, transferred to nitrocellulose membrane and probed with a HIF1α (upper panel) or β-actin (lower panel) specific antibody. The bands observed in the cadmium only WT cells are nonspecific as they are also observed in the HIF1α −/− cells. (b) BNip3 mRNA expression levels were analyzed by qRT-PCR in wild type (WT, white bars) and HIF1α −/− cells (black bars). Cells were left untreated (0), or exposed to 5 μM CdCl2 (Cd2+) or 150 μM CoCl2 (Co2+) for 24 hours. Each value was normalized to the control level in the corresponding cell line. * compared to the corresponding controls, . (c) BNIP3 protein levels were determined in wild type and HIF1α −/− cells after treatment with 150 μM CoCl2 (Co2+) or 5 μM CdCl2 (Cd2+) for 24 hours using a BNIP3 specific antibody and β-actin was used as a loading control (lower Panel).
