Fig. 8. HEF1 overproduction induces increased mRNA transcript and protein levels of downstream targets. (A) RT-PCR analysis of actin control and HEF1 targets predicted by the array analysis. (Top) Actin message was amplified from cDNA template derived from either uninduced (lanes 1-4) or HEF1 induced (lanes 5-8) HEF1.M1 cells for 23 (lanes 1 and 5), 28 (lanes 2 and 6), 33 (lanes 3 and 7) or 38 (lanes 4 and 8) cycles. M is a molecular weight marker. (Bottom) Comparative amplification of transcripts encoding MLCK, p160ROCK (ROCK), MDA7 and metalloprotease (MTLP) using normalized cDNA template derived from either uninduced (–) or HEF1 induced (+) HEF1.M1 cells. RT-PCR products shown were amplified for either 28 (MLCK, MDA7 and MTLP) or 33 (ROCK) cycles. (B) Whole-cell lysates of HEF1.M1 cells either uninduced (lanes 1, 3, 5; –) or induced (lanes 2, 4, 6; +) for 20 hours were probed with antibodies to MMP1, MMP14 or ErbB2.