FIG. 6.
p21 expression is induced in H1299-p53 cells by 12 h after the shift to the permissive temperature. H1299-p53 cells and parental H1299 cells were shifted from 39°C (p53 mutant) to 32°C (p53 wild-type). Equivalent numbers of cells were lysed at the time indicated above each lane. Total cellular protein from 2 × 105 cells was separated by SDS-PAGE and then electrophoretically transferred to nitrocellulose. p21 expression was analyzed by standard Western blotting methods. The p21/WAF-1 protein present in cellular lysates was visualized with the monoclonal antibody clone EA10 (Santa Cruz Biotechnology, Inc., Santa Cruz, Calif.).