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. 2004 Mar 1;18(5):541–558. doi: 10.1101/gad.1151204

Figure 5.

Figure 5.

Influence of Unc51.1 and SynGAP on Ras-MAP kinase activity in a reporter assay and in primary granule cells in culture. (A) A reporter assay scheme. Effects of overexpression of SynGAP with or without Unc51.1 (either wild-type or the kinase-deficient form, KR) in HEK293 cells were evaluated by measuring the Ras-MAP kinase-Elk pathway-induced luciferase reporter activation. (B) HEK293 cells were transfected with various combinations of constructs as indicated, and the relative luciferase activity was expressed as a ratio of the relative light unit (RLU) normalized against the control RLU level. Total amount of DNA used for transfection was normalized with an empty vector (pcDNA3). The data are the mean ± S.E. of reporter activities from three independent experiments. Asterisks show significant decrease in the reporter activity (p < 0.001). (C) Expression levels of the proteins were evaluated by Western blotting using myc and Flag antibodies to confirm that equivalent amount of the proteins were used in the assays. Lane numbers (lanes 3–8) correspond to column numbers in B.