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. 2012 Sep 21;7(9):e45918. doi: 10.1371/journal.pone.0045918

Figure 1. Lamin A and C are not sumoylated by sumo1.

Figure 1

(A–C) Western blots of C2C12 nuclear proteins: (A) Untransfected (UNT) or sumo1-YFP transfected, or triple transfected wild-type lamin A-CFP, DsRed2-lamin C and sumo1-YFP probed for lamin A/C. NEM was included with harvesting of selected samples to stabilize sumo conjugation. (B) Western blot A stripped and reprobed for the reversibly sumoylated protein, SP3. (C) UNT or triple transfected wild-type and mutant DsRed2-lamin C, wild-type sumo1-YFP and wild-type Ubc9-HA probed for lamin A/C. (D–E) Immunoprecipitation of endogenous and exogenous lamin A and C. (D) Nuclear extracts of sumo1-HA transfected C2C12 cells were immunoprecipitated using anti-lamin A/C antibody and western blotting was performed for lamin A/C. (E) Nuclear extracts of lamin A-CFP, lamin C-CFP, or empty CFP vector transfected COS7 cells were immunoprecipitated using anti-GFP tag antibody and western blotting was performed for the GFP tags. All protein was harvested in the presence of NEM. Emerin, a known lamin A/C binding partner, was included as a positive control for immunoprecipitation.