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. 2016 May 27;35(13):1400–1416. doi: 10.15252/embj.201593374

Figure EV2. Sufu polyubiquitylation requires interaction with the carboxyl terminus of Fbxl17.

Figure EV2

  1. Scheme representing the mapping strategy to identify Fbxl17 region required for Sufu recognition. The F‐box is highlighted in black and the leucine‐rich repeats (LRR) in yellow.
  2. Detection of Sufu and Skp1 after immunoprecipitation of Myc‐tagged Fbxl17‐truncated fragments, as indicated. An empty vector (EV) was used as a negative control. HEK293T cells were treated with MLN4924 (2 μM) for 5 h prior to collection.
  3. In vitro ubiquitylation assay of Sufu WT and Sufu mutant K257R by Fbxl17‐truncated version 318–701. All proteins were synthesized in vitro using a T7‐coupled reticulocyte lysate system and incubated for the indicated times. Arrow indicates a nonspecific band detected in reticulocyte lysates.