FIGURE 3.

CDDP has no effect on PARC and p53 interaction in chemoresistant OVCA cells. Protein A Dynabeads were incubated with PARC antibody (Bethyl, A300-096A; 2 μg/mg lysate; 1 h) and subsequently incubated for 1 h with C13* whole cell lysate (1,000 μg; lysis buffer control (LB)) obtained from CDDP treated (0–10 μm) cells for various durations. PARC pulldown and p53 content resulting from co-immunoprecipitation is unaffected by time or CDDP treatment (lanes 3–10). Also shown are controls (CTL) for potential nonspecific bands involving antibody IgG (lanes 1 and 2) and from the lysis buffer (lanes 1 and 11). To confirm the effectiveness of the CDDP treatment, PARC, p53 and GAPDH content and apoptosis counts of the above samples were examined. Results are expressed as mean ± S.E. (n = 3 replicate experiments). IP, immunoprecipitation; WB, Western blot.