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. Author manuscript; available in PMC: 2010 Feb 18.
Published in final edited form as: J Biol Chem. 2007 Jan 9;282(11):8380–8392. doi: 10.1074/jbc.M611322200

Figure 3. RSK2 induces NFAT3 activation.

Figure 3

A, RSK2 induces NFAT3 activity in a dose-dependent manner. The 3× NFAT-luciferase (Luc) reporter gene was cotransfected with pcDNA4-RSK2 into 293 cells. Following 48 h of culture, cells were disrupted with lysis buffer and analyzed for firefly luciferase activity. B, NFAT3 activity is induced by the calcium ionophore A23187. To confirm NFAT3 stimulation by A23187, the 3× NFAT-luciferase reporter plasmid was cotransfected with pcDNA3-FLAG-NFAT3 into 293 cells. Following 12 h of culture, cells were stimulated with 1 μm A23187 for 24 h and analyzed for firefly luciferase activity. C, RSK2 induces increased NFAT3 activity stimulated by A23187. To confirm that RSK2 mediates NFAT3 activity, the pcDNA3-FLAG-NFAT3 construct (20 ng) was cotransfected into 293 cells with various amounts of the pcDNA4-RSK2 construct (0, 5,10, 20, and 40 ng). Following 12 h of culture, cells were stimulated with 1 μm A23187 for 24 h and analyzed for firefly luciferase activity. For all experiments (A–C), the 3× NFAT-luciferase activity was normalized against Renilla luciferase activity (pRL-SV40), and data are expressed as the means ± S.D. of values from triplicate experiments. Also for A–C, significant differences were evaluated using Student's t test (A and B,*, p < 0.001; C,*, p < 0.05 and **, p < 0.001).