Figure 1.

A: Schematic illustration of the EMMPRIN/GFP plasmid. A 1.6-kb cDNA representing the entire EMMPRIN sequence was placed at an EcoR 1 site under the control of the CMV promoter in pcDNA 3. GFP cDNA was inserted along with an upstream CMV promoter into the EMMPRIN expression vector between NotI and XhoI sites. A polyadenylation (PA) signal was placed downstream. B: Northern blot analysis of EMMPRIN. Approximately 20 μg of total cellular RNA from plasmid alone-transfected, GFP-transfected, and EMMPRIN/GFP-transfected MDA-MB-436 breast cancer cells (from experiment 3) was size fractionated in a 1% denaturing agarose gel, transferred to a nylon membrane, and incubated with 1.7 kb of ³²P-radiolabeled EMMPRIN cDNA as a probe. Blots were analyzed by autoradiography. A single 1.7-kb mRNA transcript corresponding to the known EMMPRIN band was detected at ∼20× greater intensity in EMMPRIN/GFP-transfected cells as compared to plasmid alone or GFP-transfected cells.