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. 2009 Jan 16;284(3):1709–1717. doi: 10.1074/jbc.M807263200

FIGURE 8.

FIGURE 8.

The p115 CTF is SUMOylated in vivo. A, HeLa cells were either untransfected (lane 1), transfected with 3×FLAG-tagged CTF (lane 3), or co-transfected with plasmids encoding 3×FLAG-tagged CTF, UBC9, and HA-tagged SUMO (lanes 2 and 4). At 18 h post-transfection, cell lysates were immunoprecipitated using a monoclonal antibody to the FLAG tag coupled to Sepharose beads. The immunoprecipitated material was resolved by SDS-PAGE and subjected to Western blot analysis using monoclonal antibodies to either HA (lanes 1 and 2) or FLAG (lanes 3 and 4). Single asterisks indicate SUMOylated FLAG-tagged CTF; double asterisks correspond to FLAG-tagged CTF. The band migrating above SUMOylated FLAG-CTF (∼75 kDa) likely corresponds to a dimeric form of CTF (lanes 3 and 4). B, HeLa cells were either untransfected or transfected with HA-SUMO and treated with 5 μg/ml anisomycin for 0, 2, and 4 h. Cell lysates were analyzed by Western blot using antibodies to p115 and PARP; asterisks correspond to the cleavage products of p115 (90 kDa) and PARP (p85), respectively. C, quantitation of PARP and p115 cleavage in response to anisomycin treatment in the presence or absence of HA-SUMO expression.