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Self-SUMOylation of Arabidopsis SUMO isoforms. The Arabidopsis SUMO isoforms AtSUMO1, AtSUMO2, and AtSUMO3, each labeled with an N-terminal HA tag, were heterologously expressed, purified from E. coli, and either directly applied to the gel or preincubated with a reaction mix containing the Arabidopsis SUMO E1 and E2 enzymes. Proteins were separated on a 15% SDS-polyacrylamide gel, and SUMO mobility shifts were detected by western-blot analysis using a HA peptide-specific antiserum.
