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

Figure 3.

Figure 3

ARF promotes the sumoylation of EGR1. (A) Left panel: 293T cells cotransfected with wild-type or mutated EGR1 constructs with or without Ubc9/SUMO1 were directly lysed in NEM-RIPA buffer 48 h after transfection. Right panel: 293T cells cotransfected with Ubc9/SUMO1/EGR1 and control siRNA or p14ARF siRNA were lysed in NEM-RIPA buffer 48 h after transfection. Western blot was analysed with indicated antibodies. (B) Anti-EGR1 blot of Ni2+-NTA precipitates from ARF−/− MEFs (passage 10) transfected with Flag–EGR1 (K272R, K5R, and wild type) alone and with Flag–UBC9/His6–SUMO1 and with or without Flag–p14ARF, as indicated (upper panel); anti-Flag blot of total lysates in NEM-RIPA buffer (lower panel). (C) Anti-EGR1 blot of anti-SUMO1 immunoprecipitates from HeLa cells transfected with empty vector or Flag–p14ARF (upper panel). Lower panel: anti-EGR1 or anti-Flag immunoblot of total lysates.