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In vitro E3 ligase ubiquitin ligase activity of MBP-OsRPFs. The ubiquitination reaction contained E1, atE2s (Arabidopsis UBC10 and UBC11), maltose-binding protein-tagged OsRFPH2-2 (A), OsRFPH2-3 (B), OsRFPH2-14 (C), OsRFPH2-16 (D), OsRFPH2-23 (E), OsRFPHC-2 (F), OsRFPHC-3 (G), OsRFPHC-13 (H), or OsRFPv-6 (I), Ub, and ATP. Poly-Ub chains were detected by immunoblotting with a Ub-specific antibody.
