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Pygo2 is ubiquitylated and the extent of its ubiquitylation is decreased by the S48E mutation. FLAG-Pygo2 (A) or its Ser-48 mutant derivatives (B) was co-transfected with HA-ubiquitin, and cells were treated 24 h later with MG132, followed by immunoprecipitation using FLAG M2 beads. FLAG vector was used as a negative control. See also the legend to Fig. 7, G and H. IP, immunoprecipitation; IB, immunoblot; WCL, whole cell lysate.
