Figure 5.
Inhibition of p53/c-myc-induced apoptosis by antisense Pw1/Peg3 and a dominant negative mutant. (A) VM10 cells were transfected with vector control, Pw1/Peg3, Pw1/Peg3 antisense mRNA, and a deletion mutant Pw1/Peg3 containing amino acids 1–592 (ΔPw1). The cells were cotransfected with pEGFP as described. The transfected cells were shifted to 32°C for 24 h and analyzed by FACS as in Fig. 2. Percentages of sub-G1 cells (apoptotic) are the averages of three independent experiments. (B) Antisense Pw1 reduces Pw1/Peg3 protein levels. 293 cells were transfected with either 2 μg of Pw1/Peg3, 2 μg of ΔPW1, or the two together in different ratios (1:1 and 1:3). The HA-tagged Pw1/Peg3 was detected by immunoprecipitation-Western blotting. Transfection efficiency was monitored by cotransfecting with 0.5 μg of pEGFP.