Fig. 1
Mdm2 protein levels change in response to a high dose of UV irradiation
(A) IP-Western analysis of Mdm2 and p53. Primary Rat Embryo Fibroblasts (REF) were irradiated with a UV dose of 20 J/M2 and harvested at the indicated times following treatment. Two to three mg of cell lysate was incubated with monoclonal antibody pAb2A10 (specific for Mdm2) or monoclonal antibody pAb421 (specific for p53). The immunoprecipi-tates were electrophoresed in a SDS-Polyacrylamide gel and blotted onto a membrane. Western analysis was then performed with anti-Mdm2 serum for Mdm2 or pAb421 for p53 and 125I-protein A. (B) Quantitation of Mdm2 and p53 protein levels. The antibody-bound 125I-protein A was quantified with a Phos-pho-Imager. The relative protein level was calculated by dividing the amount of protein at each time following UV treatment by the original level with no treatment. The relative protein levels of Mdm2 and p53 were plotted in different scales as indicated.