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. 2013 Jan 24;8(1):e54282. doi: 10.1371/journal.pone.0054282

Figure 2. In vivo ubiquitination of a ZFN.

Figure 2

293T cells were transfected with pcDNA3-myc-ZFN-224 and pEFIRES-HA-ubiquitin separately and together. ZFN ubiquitination was confirmed by co-immunoprecipitation with an anti-myc antibody and immunoblotting with an anti-HA antibody. To cross confirm, co-immunoprecipitation was performed using anti-HA antibody and immunoblotting with an anti-myc antibody. MG132 treatment further increased ZFN ubiquitination, as shown by the increased amounts of the high molecular weight smear, which represents polyubiquitin coupled with the ZFN.