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Fig. 8. ShRNA expression interferes with Hint1 expression and enhances TCF–ß-catenin transcriptional activity. (A) Lysates from HeLa cells stably transfected with interfering shRNA-Hint1430/431 or non-functional shRNA-Hint1432/433 were analyzed for Hint1 expression by western blotting with anti-Hint1 ({alpha}-21) antibody. (B) Increasing amounts of interfering shRNA-Hint1430/431 increase TCF–ß-catenin-mediated transcription whereas control shRNA-Hint1432/433 did not show an effect. By contrast, shRNA directed against ß-catenin reduces reporter gene activity. When HeLa cells stably transfected with control shRNA-Hint1432/433 were transiently transfected with interfering shRNA-Hint1430/431 again transcriptional activity was increased whereas vice versa no effect was detectable. (C) Quantitative RT-PCR analysis of Axin2 expression in HeLa cells transiently transfected with Hint1 or interfering shRNA-Hint1430/431.





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