Fig. 7.
Regulation of p21waf1 mRNA in cyclin K–expressing cell lines.
(A) NIH3T3 cells were stably transfected with an expression vector encoding cyclin K (lane 1) or the corresponding parental plasmid (lane 2). Following G418 selection, overexpression was verified using nuclear extracts analyzed by Western blot with the M2 Flag monoclonal antibody. (B) NIH3T3 cells were serum starved for 2 days and stimulated for 2 hours with IL-6 (10 ng/mL; lanes 1, 4) or left untreated (lanes 2-3). Total RNA was prepared and 10 μg RNA was subjected to Northern blot analysis using a mouse cDNA probe. The membrane was striped and reprobed with a 18S oligonucleotide (bottom panel).
