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. 2017 Apr 3;36(31):4405–4414. doi: 10.1038/onc.2017.69

Figure 3.

Figure 3

SPZ1 activates the TWIST1 promoter through its G-box. (a) Forced SPZ1 (SPZ1-GFP) activated EMT and stem cell-like marker expression in Hep G2 and Hep 3B cells. Arrow: position of indicated protein. (b) The expression of SPZ1, TWIST1 mRNA was examined in Tet-ON inducible SPZ1-GFP transformants until 8 h after addition of DOX. P<0.001. (c) Western blotting of induced SPZ1 and TWIST until 8 h after induction with DOX is shown. (d) Schematic presentation of a series of luciferase deletions of TWIST1 promoter. Relative luciferase activity in SK-Hep1 was calculated as described in the Materials and methods section. P<0.001. (e) Relative DNA-binding activities in SK-Hep1 were measured as described in the Materials and Methods section. P<0.001. (f) The transactivation activity of SPZ1 on TWIST1 promoter was determined as described in the Materials and Methods section. a, P<0.001. (g) ChIP assay of the TWIST1 promoter with SPZ1 antibodies in various hepatoma lines as described in the Materials and Methods section. a, P<0.001. # indicates cell lines with a lower expression of SPZ1.