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Figure 6


Fig. 6. Effects of a dominant negative TCF4 on RAS-induced activation of ERK and ELK1 reporters. (A) DLD-1 cells were grown and transfected with 0.1 µg of empty pMT3 vector or 0.1 µg of pMT3 RAS-L61. Where required, they were co-transfected with 0.5 µg of the MYC-tagged dnTCF4 expression vector, {Delta}N-TCF4E (Tetsu and McCormick, 1999). 48 hours after transfection, cell extracts were prepared, and p-ERK, RAS-L61, {Delta}N-TCF4E (dn-TCF4-MYC), ERK and {alpha}-tubulin proteins were detected by western blotting. Anti-MYC antibody was used to detect dn-TCF4-MYC. (B) DLD-1 cells were grown and transfected with 0.1 µg of empty pMT3 vector or 0.1 µg of pMT3 RAS-L61. Where required, they were co-transfected with 0.5 µg of MYC-tagged dn-TCF4 expression vector, {Delta}N-TCF4E. 0.5 µg of ELK1 trans-reporter was co-transfected, as described in Fig. 1B. Extracts were prepared 48 hours after transfection, and luciferase activities measured. Each data point represents the average of three independent analyses.





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