Fig. 4

ZFX regulates the transcription of WNT3. A Chromatin immunoprecipitation (ChIP) was performed to analyze the interaction between ZFX protein and the WNT3 gene. A schematic graph is displayed to show the primer sets designed for ChIP analysis (upper panel). The results of ChIP–qPCR analysis with various primer sets are shown (lower panel). B The promoter region of WNT3 (−1137 to +164) was subcloned into a reporter vector (designated pGL3–FL), and this vector was transfected into ZFX-silenced and control (scramble) K562 cells. The relative luciferase activity of this reporter is shown. C pGL3–FL contained multiple putative ZFX binding sites; therefore, various deletion mutants were generated. Then, the activities of these vectors were assessed in K562 cells (n = 3). D One putative ZFX binding site (−106 to −95) was deleted to generate pGL3–Mu, and its activity was compared with that of pGL3#6 and pGL3#7 (n = 4), respectively. Data are presented as the mean ± SEM, and Student’s t-test was used to estimate the P-values (*P < 0.05, **P < 0.01, and ****P < 0.0001)