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. 2017 Sep 15;18(9):1986. doi: 10.3390/ijms18091986

Figure 5.

Figure 5

Hsp90α facilitated the nuclear translocation of c-Myc and EZH2 to BMI1 promoter. (A) The binding of EZH2 or Hsp90α on the E-Box region of BMI1 promoter in MDA-MB-231 mammospheres was determined by the ChIP method with anti-EZH2 or anti-Hsp90α antibody, respectively. Data were presented as the relative expression level to input chromatin of each sample. * p < 0.05; ** p < 0.01; (B) The purity of the cytoplasmic or nuclear fraction from MDA-MB-231 mammospheres was confirmed by Western blot analysis of tubulin (cytoplasmic specific protein) or histone 1 (nuclei specific protein). An equal amount of nuclear or cytoplasmic proteins from DMSO- or 17-DMAG-treated MDA-MB-231 mammospheres was used for immunoprecipitation analysis; (C) The contents of Hsp90/ EZH2/c-Myc complex in cytoplasmic or nuclear fraction proteins of MDA-MB-231 mammospheres were determined by immunoprecipitation with anti-Hsp90α antibody and analyzed by Western blot. N, nuclear; C, cytoplasmic; B, beads only; IP, immunoprecipitation.