Fig. 4: Phosphorylated and non-phosphorylated FTZ-F1 preferentially bind to distinct DNA motifs and both function in the nucleus.

a, b EMSA assay validates that the non-phosphorylated FTZ-F1^T288A(-P) (a) and the phosphorylated FTZ-F1^T288D(P) (b) specifically bind to the FBS (a), and FBS^P (b) respectively. The concentrations of the wild-type and mutant probes were 20 fmol; the concentrations of competing cold probes were 100 and 500 fmol. The mutant sequences were 5′-CGCACACACGT−3′ for FBS and 5′-GGCTCCGAAC-3′ for FBS^P. c Y1H assays verifying the direct binding of non-phosphorylated FTZ-F1^T288A(-P) to FBS, and the direct binding of phosphorylated FTZ-F1^T288D(P) to the FBS^P using wild-type or mutated binding sites as described below. EV empty vector; positive control, pGADT7-p53 + pABAi-p53. d Subcellular localization of non-phosphorylated FTZ-F1^T288A(-P) protein and the phosphorylated FTZ-F1^T288D(P) protein. The nuclei were stained with DAPI, the Sf9 cells or cells transfected with the empty plasmids (Pie-EGFP-N1) were used as controls. Scale bar: 10 μm. Source data are provided as a Source Data file.