Figure 1.

Characterization of MEF^RAS7A+ and MEF^RAS7A- cells. (A) MEF^RAS7A+ cells were generated from a mouse embryonic fibroblast cell line carrying a floxed Atp7a allele and transformed by sequential transfection of SV40 large T-antigen and mutant H-RAS oncogene. Isogenic MEF^RAS7A- cells were generated by infecting with adenovirus carrying the Cre recombinase. (B) Immunoblot analyses. Cell lysates were separated by SDS-PAGE and immunoblotted with the indicated antibodies. (C) Copper concentration determination within cultured MEF^RAS7A+ and MEF^RAS7A- cells by inductively coupled plasma mass spectrometry (mean ± S.E.M; ***P<0.001; n = 3). (D) Cell proliferation was evaluated using WST-1 colorimetric assay by measuring the absorbance at 440nm (mean ± S.E.M; n = 3). (E) Representative images of MEF^RAS7A+ and MEF^RAS7A- cells Scale bars: 50µM. (F) Cell size determination of MEF^RAS7A+ and MEF^RAS7A- cells (mean ± S.E.M; n = 30 cells in three fields).