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. 2014 Mar 20;9(3):e92746. doi: 10.1371/journal.pone.0092746

Figure 5. Nuclear localization is essential for ATBF1 to undergo SUMOylation.

Figure 5

A. Mutation of the NLS at residues KRK2615–2617 had no effect on the SUMOylation of ATBF1 fragment 8 in the in vitro assay, as detected by IB. B. Failure in nuclear localization prevents the SUMOylation of ATBF1. HA-tagged wildtype ATBF1 (HA-ATBF1) or a mutant that was deficient in the NLS (HA-ATBF1-NLSm) were co-transfected with GFP-tagged SUMO1 into 22Rv1 cells, and cell lysates were pulled down with anti-HA beads, and blotted with anti-GFP (upper) or anti-ATBF1 antibody (lower) to detect SUMOylated ATBF1. The native ATBF1 protein is about 400 kD.