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. Author manuscript; available in PMC: 2009 Apr 1.
Published in final edited form as: Mol Cancer Ther. 2008 Apr;7(4):905–914. doi: 10.1158/1535-7163.MCT-07-0515

Figure 6.

Figure 6

Effects of UBE1L on ISG15ylation of the PML domain of PML/RARα. BEAS-2B cells were individually transfected with the PML domain fragment of PML/RARα containing expression construct. Indicated groups were cotransfected with or without UBE1L. Following transfection, respective groups were treated with either ALLN (100 μmol/L) or vehicle (DMSO) for 4 (A and B) or 6 (C and D) additional hours. Immunoprecipitation followed by immunoblot analyses were done as described in Materials and Methods. The filters were probed with either a monoclonal anti-HA antibody to detect PML (A and C) or a monoclonal anti-ISG15 antibody to detect ISG15ylated forms of PML (B and D). Open arrows, positions of unconjugated PML protein; closed arrows, positions of post-translational modification products; Control, whole-cell lysates of COS-1 cells transfected with PML serve as a signal for the unconjugated form of PML. These experiments established that UBE1L confers ISG15 conjugation to the PML domain of PML/RARα with subsequent repression of this protein.