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. Author manuscript; available in PMC: 2016 Jun 15.
Published in final edited form as: Prostate. 2015 Mar 1;75(9):907–916. doi: 10.1002/pros.22974

Figure 3. SLUG regulates PTEN promoter activity through direct interaction.

Figure 3

(A) Diagram of PTEN promoter-driven luciferase reporter. The luciferase reporter (PTEN-Luc) driven by human PTEN promoter (left panel) contains two E-box sites (E-box1 and E-box2).

(B) PTEN Overexpression of SLUG suppresses PTEN promoter activity. PTEN-Luc and an increasing dose of Slug expression vector was transfected together into 293T (left panel) and 22RV1 (right panel) cells. pCMV-LacZ was included into the transfections as a control. Data were normalized by pCMV-LacZ activity, and graphed as fold change. ** p < 0.01.

(C) ChIP assay for SLUG binding to PTEN promoter. Retroviruses expressing Flag-tagged SLUG (Slug-Flag) were infected into DU145 and 22RV1 cells. DNA sequences that are associated with SLUG were precipitated with anti-Flag antibody. Anti-HA antibody was included as a negative control. Using specific primers, the precipitated DNAs containing the PTEN promoter were then detected by PCR (left panel) and qPCR (right panel). ** p < 0.01.