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. Author manuscript; available in PMC: 2009 Oct 27.
Published in final edited form as: Cancer Res. 2008 Sep 15;68(18):7650–7660. doi: 10.1158/0008-5472.CAN-08-1137

Figure 3. Cdk3 enhances transactivation activity of ATF1.

Figure 3

(A) A pGal4-ATF1 expression vector was constructed as indicated (left panel). The transactivation activity of ATF1 was measured in T98G cells transfected with pGal4-ATF1 or pGal4-ATF1 S63A as described in “Materials and Methods” (middle panel). The mammalian two-hybrid assay was performed in HEK293 cells to assess the interaction of pBIND cdk3 with pACT-ATF1 or pACT-ATF1 S63A (right panel). B, C, A c-jun-luciferase reporter plasmid (B) or a c-fos-luciferase reporter plasmid (C) was co-transfected with increasing amounts of a pcDNA4-ATF1 plasmid into T98G cells and then firefly luciferase activity was analyzed (left panels). The pcDNA4-ATF1 and c-jun-luciferase or c-fos-luciferase reporter plasmid were co-transfected with increasing amounts of pCMV-cdk3 into T98G cells and then firefly luciferase activity was analyzed (right panels). For all experiments (A-C), the firefly luciferase activity was analyzed at 36 h and normalized against the Renilla luciferase activity. Data are shown as the mean ± S.D. of values obtained from triplicate experiments. Significant differences were evaluated using the Student’s t–test (*, p < 0.05; **, p < 0.001).