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. 2016 Aug 16;291(41):21584–21595. doi: 10.1074/jbc.M116.730978

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — cAMP-binding site within PDZ-GEF1 is required for cAMP activation of Ras. A, Hek293 cells were transfected with NS shRNA or shRNA against PDZ-GEF1 and either vector or FLAG-tagged shRNA-resistant wild type PDZ-GEF1 (WT*). Cells were treated with F/I for the times indicated. The levels of pERK and total ERK2 are shown in the 1st and 2nd panels, respectively. The levels of PDZ-GEF1 are shown in the 3rd panel. B, cells transfected with NS shRNA or shRNA against PDZ-GEF1 and either vector, FLAG-tagged shRNA-resistant wild type PDZ-GEF1 (WT*), or FLAG-tagged shRNA-resistant PDZ-GEF1 K211D mutant (K211D*). Cells were treated with F/I for the times indicated. The levels of Ras-GTP are shown in the 1st panel. The levels of pERK, total Ras, total ERK2, and FLAG-PDZ-GEF1 are shown in the lower four panels, respectively.