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. 2015 Dec 2;7(3):3283–3296. doi: 10.18632/oncotarget.6488

Figure 3. The interference of REIC/DKK-3 with SGTA-SGTA dimerization.

Figure 3

A. The interference of REIC/DKK-3 with SGTASGTA dimerization was analyzed in a mammalian two-hybrid assay. Two-hybrid constructs of full-length SGTA (GAL4 DBD) and fulllength SGTA (VP16 AD) were co-transfected into 293T cells. HA-tagged full-length REIC/DKK-3 was also transfected under the indicated conditions. Luciferase activity was measured to determine the extent of SGTA-SGTA dimerization; the results are shown in the bar graphs. The data are from four independent experiments. The bar graphs show the luciferase activities on the interaction of SGTA-DBD, SGTA-AD and HA-tagged empty vector column as 100%. *, A significant difference was observed. B. For the pull-down (PD) assay, the full-length human SGTA cDNA was cloned into both the pFN21A and pMACS Kk.HA-C plasmid vectors. A pDNR-CMV vector encoding the fulllength REIC/DKK-3 was also used. These plasmids were co-transfected, and the cell lysates from Halo-tagged SGTA- and/or HA-tagged SGTA- and/or REIC/DKK-3-transfected 293T cells were analyzed. The samples pulled down by Halo-tagged SGTA were analyzed by Western blotting (WB) with the indicated antibodies.