A: Southern blot analysis of normal mucosa (N) and their seven corresponding cases of colonic adenocarcinomas (T1–T7), cases No. 1, 2, 4, and 5 are poorly differentiated whereas cases No. 3, 6, and 7 are moderately differentiated. Genomic DNA was digested with BglII, fractionated by electrophoresis in agarose gel, transferred onto membranes and hybridized with PRAD1 and β-actin. Tumors number 1–6 (Lanes 1–6) show different degrees of PRAD1/cyclin D1 amplification, tumor number 7 (lane 7) was not amplified. B: Southern blot analysis of 3 cases of adenocarcinomas (T) and matched normal colonic mucosa (N). Genomic DNA was digested with EcoRI, fractionated by electrophoresis in agarose gel, transferred onto membranes and hybridized with PRAD1 and β-actin probes for loading control. The identification of the 3 tumors is the same as in Fig. 3A with amplification of PRAD1/cyclin D1 in tumors number 4, 5 (Lanes 1, 2) but not 7 (Lane 3).