FIG. 2.

Coimmunoprecipitation of MDM2 with p53. p53 was immunoprecipitated, with the CM5 antibody, from lysates of GM00011 cells carrying the retroviral vector alone or expressing E7. Following transfer to nitrocellulose, p53 and MDM2 were detected with antibodies DO-1 and 2A10, respectively. Lane 1, 3 μl of a rabbit reticulocyte lysate programmed with p53 RNA and 3 μl of a rabbit reticulocyte lysate programmed with RNA from human mdm2; lane 2, 150 μg of total lysate from NDFs carrying the vector alone; lane 3, immunoprecipitate (IP) from 150 μg of lysate from NDFs carrying the vector alone; lane 4, immunoprecipitate from 300 μg of lysate from NDFs carrying the vector alone; lane 5, immunoprecipitate from 900 μg of lysate from NDFs carrying the vector alone; lane 6, 150 μg of total lysate from NDFs expressing E7; lane 7, immunoprecipitate from 15 μg of lysate from NDFs expressing E7; lane 8, immunoprecipitate from 75 μg of lysate from NDFs expressing E7; lane 9, immunoprecipitate from 150 μg of lysate from NDFs expressing E7; lane 10, immunoprecipitate from 300 μg of lysate from NDFs expressing E7; lane 11, immunoprecipitate from 450 μg of lysate from NDFs expressing E7; lane 12, immunoprecipitate from 900 μg of lysate from NDFs expressing E7.