Fig. 2.

Affinity purification of SUMOylated proteins from Arabidopsis. Crude extracts (Cr) from wild-type (WT) and the His-H89R-SUM1 sum1-1 sum2-1 seedlings were subjected to sequential Ni-NTA (Ni-1), anti-SUMO1 (@SUMO), and Ni-NTA affinity chromatography (Ni-2). The flow through (FT), eluate (E1-3), and solubilized E1 (ES) fractions were separated by SDS-PAGE and either stained for total protein (Left) or subjected to immunoblot analysis with anti-SUMO1 antibodies (Right). Immunoblot loads were proportionally adjusted to allow direct comparisons among samples. A longer exposure of the immunoblot is included to show the levels of His-H89R-SUMO1. ▸, free His-H89R-SUMO1. *, large subunit of RUBISCO.