Zinc, cobalt, and cadmium induce MTF-1 translocation into the nucleus and increased MT-1 mRNA. (A) NIH3T3 cells were transfected with pcDNA-mouse-MTF-1-Flag. Two days later, the cells were exposed to the specified transition metals (100μM) for 2 hours. Cells were fixed and MTF1-Flag localization detected by indirect immunofluorescence as described in “Immunofluorescence and Microscopy.” The right hand panel is a quantification of the imaging where > 200 cells were examined for nuclear or cytosolic MTF-1-Flag. (B) Macrophages were exposed for 16 hours to serial concentrations of transition metals ranging from 0 to 200μM, total RNA was harvested and semiquantitative RT-PCR were performed for MT-1 and actin mRNA. All experiments were performed a minimum of 3 times.