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. 2006 May 18;25(11):2358–2367. doi: 10.1038/sj.emboj.7601149

Figure 1.

Figure 1

Negative regulation of IFN signaling by Ubp43 is independent of its enzymatic activity. (A) Total protein extracts were prepared from ubp43+/+ and ubp43−/− MEFs or ubp43−/− MEFs stably expressing Ube1L siRNA, wt murine Ubp43, or Ubp43C61S after mIFN-β (1000 U/ml) treatment for 0′, 30′, and 10 h and analyzed by Western blotting with the respective antibodies. (B) 293T cells were transiently transfected with vector-control (lane 1) or plasmids containing 6xHis-ISG15, HA-Ube1L, and Flag-Ubc8 in the absence (lane 2) or the presence of wt (lane 3) or mutant (c61s) Ubp43-V5 (lane 4). The level of ISG15 conjugation was determined by Western blotting with anti-mISG15 antibodies. Blots were stripped and reprobed with anti-HA, anti-Flag, or anti-V5 antibodies to ensure equal levels of protein expression.