Figure 5. Time course stability of CYP1B1 mutant polypeptides found in Spanish PCG patients. cDNA constructs encoding the different mutations were transiently expressed in HEK-293T cells. Cells were treated with cycloheximide, a protein synthesis inhibitor, and the amount of the different CYP1B1 polypeptides, tagged with the myc epitope at their COOH-terminal ends, were determined by densitometry of the signals detected in western blots using an anti-myc monoclonal antibody (Santa Cruz) at the indicated time points. Transfection efficiency was assessed by co-transfection with a cDNA construct encoding GFP. When required, CYP1B1 protein levels were corrected for transfection efficiency. Relative amounts of CYP1B1 are expressed as a percentage of levels at time 0 h. Error bars represent the SEM of triplicate experiments.