Fig. 2.
The SC motifs in GR are sites for SUMO modification. COS-7 cells were transfected with vectors for WT (p6RGR) or SC mutant (p6RGR K297R/K313R) GR- and pCDNA3-based vectors for HA-tagged SUMO-1, -2, and -3. After a 16-h incubation with either vehicle or Dex, GR immunoprecipitates were resolved by SDS/PAGE and processed for immunoblotting by using anti-HA (A) or BuGR (B) antibodies. (C) In vitro SUMOylation reactions were performed as described in Methods by using in vitro-transcribed and -translated [35S]-labeled WT or SC mutant (K297E/K313E) GR as substrates in the presence of the indicated components.