Figure 5. KDM4B interacts with BMYB and activates BMYB-targeted PLK1 transcription.
(A) PLK1-luc reporter activities in cells transfected with full-length (1.5 kb), deletion (1.0, 0.5 kb) or point-mutant (x) of a PLK1-promoter construct and a vector that expresses KDM4B, KDM4BH189G, or BMYB as indicated. (B) PLK1-luc activity in control, KDM4A (2A), or KDM4B (2B) knocked down cells that were transfected with various amount of BMYB. (C) LNCaP cell lysates were immunoprecipitated with control IgG or anti-KDM4B antibody, respectively. Immunoprecipitates were Western blotted with antibodies against BMYB and KDM4B. (D) 293T cells were transfected with Flag-BMYB and HA-KDM4B (left lower panel) or various deletion mutants of KDM4B (right panel). Whole cell lysates were immunoprecipitated with anti-Flag antibody and immunoprecipitates were Western blotted with anti-HA antibody. (E) ChIP-qPCR assays showing the relative occupancy of H3K9me3, KDM4B, BMYB, H3K36me3, and H3K4me3 at the PLK1 promoter in LNCaP cells treated with compound B3 (1 μM) or vehicle DMSO. N=3, mean ± SD. *, p<0.05.
