Fig. 1

UXT interacts with EZH2. a Identification of the interaction between UXT and C­terminal fragment of EZH2 by the yeast two­hybrid approach. b EZH2 interacts with UXT in a yeast two­hybrid assay. Yeast AH109 cells were co­transformed with the indicated combinations of plasmids (left, top). The yeast colonies containing these plasmids were grown on SD-Leu-Trp (left, bottom) agar plates and on SD-Leu-Trp-His with 25 mM 3AT (3­Amimo­1,2,4­Triazole) agar plates (right, top) and were tested by the X­Gal assay (right, bottom). Abbreviations: AD, pGADT7; BD, pGBKT7. c In vitro GST pull-down assays. The GST, GST-EZH2, or GST-UXT purified from E. coli was incubated with Flag­UXT- or Flag-EZH2­expressing HEK293T lysates and precipitated with glutathione­Sepharose, respectively. Precipitates were subjected to SDS­PAGE and examined by immunoblotting with anti­Flag or anti-GST antibodies. d Co­IP of UXT and EZH2. Immunoprecipitation of Myc–UXT (left) and HA-EZH2 (middle and right) with anti-FLAG antibody, respectively. Cell extracts from 293 T cells (left and middle) or 786-O cells (right) with equal amounts of proteins were immunoprecipitated with an anti-FLAG antibody and analyzed by Western blotting using anti-Myc or anti-HA antibodies