Figure 10.

SPF1 desumoylates sumoylated EDA9 proteins in vivo. 35S_pro:EDA9:YFP was expressed in Columbia (Col) wild-type plants and introduced into spf1-1 and spf1-2 mutants by crossing, and then homozygous plants were used for western blots. A, EDA9:YFP proteins were purified from 35S_pro:EDA9:YFP seedlings by GFP antibody affinity beads and then applied to western blots, which were probed by GFP and SUMO antibodies (Murtas et al., 2003). Each lane was loaded with 20 µL of the eluate from the GFP beads. B, Whole proteins were extracted from the transgenic seedlings and then applied to western blots, which were probed by GFP and ACT antibodies. ACT was used as a loading control. The top band is predicted as sumoylated EDA9:GFP (SUMO:EDA9:GFP), as shown in A. C, Siliques of 35S_pro:EDA9:YFP Col and 35S_pro:EDA9:YFP spf1-2. The experiments were repeated at least three times. Bar = 200 μm.