Figure 5.

NEP complexes with the p85 subunit of PI3-K. (a) 500 μg of total cell lysates were immunoprecipitated with anti-p85 antibody, anti-NEP antibody J5 or control antibody, separated by SDS-PAGE, transferred to nitrocellulose, and Western blotted with anti-NEP antibody 5B5 or anti-p85 antibody. (b) Cell lysates derived from WT-5, TN-12, and M-22 cells cultured with (+) and without (–) 1 μg tetracycline were immunoprecipitated with anti-p85 antibody, anti-NEP antibody J5 or control antibody and analyzed as described in a. Total cell lysate (TCL) from LNCaP cells was positive control.