FIG. 7.

Reduced Wnt signaling in cFLIP-depleted cells. (A) A549 cells in 60-mm-diameter dishes were transfected with a total of 2.4 μg of DNA containing 1.2 μg of control shRNA vector (open bars) or FLIP shRNA vector (solid bars), TOP-TK-Luc (400 ng), and pRL-TK (80 ng) for 12 h and replated in 48-well plates. After 24 h, the cells were treated with conditioned medium containing 500 ng of Wnt3a/ml for 12 h. The luciferase activity was measured and is expressed as the increase (n-fold) compared with the level observed in control cells treated without Wnt3a. The data represent means of triplicate determinations. The error bars indicate standard deviations. (B) Cell lysates from A549 cells that had been transfected with the indicated shRNA vectors were analyzed by Western blotting with anti-FLIP (top) and antitubulin (bottom).