Figure 7.

mRNA expression of MAZ in normal human liver, in peritumoral liver, in liver cirrhosis, in HCC and in CCC, MAZ-binding to the Prox1 promoter. (A) mRNA expression levels of MAZ in normal human liver, in non-cirrhotic peritumoral liver, in liver cirrhosis, in cirrhotic peritumoral liver, in HCC and in CCC compared to the average expression of normal liver, which is displayed as 1. HCC displayed a significant upregulation, marked with asterisk. The same individual samples are marked as the ones on Fig. 5. (B) EMSA reaction of cell nuclear extracts of HepG2 (2), Hep3B (3) and Mz-Cha2 (4), cell lines, and of nuclear extracts of HCC (5–6) and CCC samples (7) incubated for 30 minutes with the MAZ-consensus binding oligo-containing 5'-flanking sequence of the "Prox1-7902"-transcript (Fig. 1) (left panel). When the nuclear extracts were reacted with the same radiolabelled consensus double-stranded oligo in the presence of a 100-fold excess of an unlabeled MAZ-consensus oligo (published in ref. [23]) the intensity of mobility shift significantly decreased (right panel); sample 1: negative control, without nuclear extract. (C) EMSA reaction of cell nuclear extracts of Hep3B (2–3) cells incubated for overnight with the MAZ-consensus binding oligo-containing 5'-flanking sequence of the "Prox1-7902"-transcript, without (2) and with (3) anti-MAZ antibody; sample 1: negative control, without nuclear extract. The strong unspecific band might represent other transcription factors binding to the same sequence. In the presence of the anti-MAZ antibody a supershift band was observed (lane 3).