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. 2012 Nov 30;7(11):e50637. doi: 10.1371/journal.pone.0050637

Figure 9. Lack of SUMOylation increases the protein-protein interaction between FOG-2 and GATA-4.

Figure 9

COS-7 cells were transfected with constructs containing GFP alone, GFP-FOG-2 wt and 4KR mutant, HA-SUMO-1 and GATA-4 as indicated in the figure. Cell lysates were obtained in the presence of NEM. (A) Immuno-precipitation experiments were performed in cell extracts using magnetic beads coated with an anti-GFP antibody. Immuno-precipitated complexes were resolved by SDS-PAGE and blotted with anti-FOG-2 or anti-GATA-4 antibodies. (B) Cell lysates (5% input) were resolved by SDS-PAGE and blotted with anti-FOG-2 or anti-GATA-4 antibodies. Note that FOG-2 is SUMOylated by endogenous SUMO when GATA-4 is co-expressed (lane 2, upper panels). (C) The immuno-precipitation was repeated and GATA-4 was quantitated by densitometry using ImageQuant TL 1D, version 7.0 (GE Healthcare). The graph shows GATA-4 enrichment relative to immuno-precipitated FOG-2 (percentage).