Figure 6

Gene expression in LT97 adenoma cells overexpressing COX-2. LT97 cells were infected with an adenoviral vector expressing COX-2. Control groups were infected with control viruses containing no ectopic gene and cells exposed to the COX-2 inhibitor SC236. (A) Cells were homogenised in RIPA buffer and COX-2 protein analysed by Western blotting. (B) Prostaglandin E₂ secretion into the medium was determined by ELISA. (C) RNA was isolated from control transfectants and COX-2-LT97 cells in the presence and absence of the COX-2 inhibitor SC236 and c-fos expression was analysed by standard RT–PCR. Polymerase chain reaction products were seperated on 6% acrylamide gels and band intensities were semi-quantified using the Image Quant software. The results represent the mean±s.e.m. from three independent experiments. ^*Increased above control at P<0.05. (D) c-jun in nuclear extracts from control cultures and COX-2-LT97 cells in the presence and absence of SC236 was quantified as described in Figure 2.