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. 2008 Dec 4;28(1):21–33. doi: 10.1038/emboj.2008.238

Figure 4.

Figure 4

Sumoylation of EGR1 is required for PTEN induction. (A) Anti-EGR1 blot of 293T cells transfected with Flag–EGR1 alone or with Ubc9/SUMO1 and p14ARF as indicated (upper panel). Anti-Flag (second panel), anti-PTEN (third panel), and anti-β-actin (bottom panel) blots of the same experiment. (B) Activation of the PTEN-luc reporter by WT–EGR1 and by K272R–EGR1 in the presence of Ubc9/SUMO1 or Ubc9 siRNA. The 293T cells were transfected with PTEN-luc and SUMO1/Ubc9 or control siRNA or Ubc9 siRNA, and empty vector cDNA3, or WT-EGR1, or K272R-EGR1, as indicated. Luciferase activity was measured 24 h later. (C) Left panel: immunoblots for PTEN, EGR1, p19ARF, β-actin, Akt1, and phospho-Akt1 from ARF−/− (passage 12) and ARF+/+ (passage 5) MEFs. Right panel: anti-EGR1 blot of anti-SUMO1 immunoprecipitates from the same lysates.