Dose-dependent SUFU repression of GLI activity in NIH3T3 cells. A

<p>. Western blot analysis of extracts from NIH3T3 cells, transfected with expression constructs for the SUFU variants, and detected with a SUFU antibody. Due to the partial co-migration with endogenous mouse Sufu, the SUFU-FL expression in the right lane was calculated by subtracting from the total signal the mouse Sufu signal. This was determined based on the Sufu expression in the left lane following normalization with the tubulin internal control. Note that the quantitation revealed that the level of SUFU-FL (right lane, upper protein band) is 188% the level of SUFU-ΔC (middle lane, lower protein band). <b>B–E</b>, Different amounts of Myc-tagged SUFU-FL or SUFU-ΔC expression constructs were co-transfected with 50 ng GLI expression constructs (Panel <b>B</b>, GLI1FL; panel <b>C</b>, GLI1ΔN; panel <b>D</b>, GLI2FL; panel <b>E</b>, GLI2ΔN), 12xGLIBS-luc and pRL-SV reporter plasmids, and the luciferase activity was measured. Error bars indicate the standard deviation.