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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 6. — Small G protein Rap1 mediates the sustained portion of ERK activation by cAMP. A, Hek293 cells transfected with either NS or shRNA against Rap1a/b 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 efficacy of the Rap1 knockdown is shown in the 3rd panel. B, cells transfected with either NS shRNA or shRNA against Rap1a/b were treated with isoproterenol (Iso) for the times indicated. The levels of pERK and total ERK2 are shown in the 1st and 2nd panels, respectively. The efficacy of the Rap1 knockdown is shown in the 3rd panel. C, cells were transfected with NS shRNA or shRNA against human Rap1b and either vector or FLAG-tagged shRNA-resistant Rap1b (bovine FLAG-Rap1b). Cells were treated with F/I for 10 min or left untreated, lysed, and examined by Western blotting. The levels of activation of ERK (pERK) and levels of total ERK2 are shown in the 1st and 2nd panels, respectively. The levels of FLAG-Rap1b (bovine) and endogenous (endo) human Rap1 are shown in the bottom two panels, respectively.