Figure 6From: p8 inhibits the growth of human pancreatic cancer cells and its expression is induced through pathways involved in growth inhibition and repressed by factors promoting cell growthp8 transcription is repressed by the Raf→MEK→ERK pathway. Panc-1 and BxPc-3 were cultivated in 30 mm diameter culture dishes for 24 hours and then transiently transfected with 0.5 μg of p8-CAT and 0.5 μg of pCMV/βgal plasmids. Expression plasmids pcDNA RAF BXB encoding the wild-type RAF, pcDNA RAF 301 K375W encoding a dominant negative RAF, and pcDNAIII HA ERK2 encoding wild-type ERK2 (0.5 μg) were co-transfected with p8-CAT and pCMV/βgal plasmids as indicated. Cell extracts were prepared 24 hours after transfection and CAT and β-galactosidase activities were measured. CAT activity was normalized to β-galactosidase activity. Experiments were carried out in triplicate and repeated two or three times.Back to article page