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. 2008 Jul 9;13(8b):1719–1727. doi: 10.1111/j.1582-4934.2008.00423.x

Figure 5.

Figure 5

Repression of Siah1 expression attenuates E2F1‐repressed β‐catenin/TCF activity. (A) H1299 cells were transfected with gene‐specific shRNA construct pSuper‐Siah1 (1ug) or pSuper empty vector (1ug). Forty‐eight hours after transfection, cells were harvested for preparation of total RNA and semi‐quantitative RT‐PCR analysis with primer pairs specific to Siah1 and β‐actin. Actin (control) was then performed. (B) H1299 cells were transfected with either Siah1 shRNA construct pSuper‐Siah1 (200 ng) or pSuper empty vector (200 ng). Forty‐eight hours after transfection, 200 ng of pTop‐flash and 5 ng of pRL‐CMV were further introduced into each transfected cells. An additional 24 hrs later, cells were harvested for measuring dual luciferase activity as described in Fig. 2. Luciferase values were corrected for transfection efficiency with Renilla activity. All the experiments were independently performed in triplicate. The data of triplicate experiments and their average were indicated in the figure. Error bars indicate S.D. (standard deviation). ***P < 0.001; **P < 0.01.