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Fig. 8. Combinatorial effect of PITX2, LEF-1 and ß-catenin on LEF-1 promoter activity. (A) CHO cells were transfected as in Fig. 1, with CMV-PITX2C, CMV-LEF-1 or CMV-ß-catenin S37A, or combinations of each, and the CMV empty expression vector. The activities are shown as mean fold activation compared with the LEF-1 promoter plasmid without protein expression and normalized to ß-galactosidase activity (± s.e.m. from four independent experiments). (B) Expression of PITX2 in transfected CHO cell lysates; approximately 10 µg of lysate was used in the western blot. PITX2 expression was similar in cells transfected with LEF-1 or ß-catenin, or both. Bacteria expressed PITX2 protein (100 ng) was used as a control; the protein expressed in transfected cells migrates slower than the bacterially purified protein due to the presence of a Myc/His C-terminal tag.
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