(A) SUMOylation of FLAG-Atxn1 82Q in HeLa cells in the absence or presence of HA-PML cells, and without or with MG132 treatment. The amount of Atxn1 82Q DNA used for transfected were adjusted to yield comparable levels of the unmodified protein.
(B and C) SUMOylation of FLAG-Atxn1 82Q (B) and FLAG-nFlucDM-GFP (C) in control and PML siRNA-transfected HeLa cells treated without or with MG132.
(D and E) SUMOylation of purified HA-Atxn1 82Q-FLAG was performed in the presence of recombinant FLAG-PML, FLAG-PML M6, and SUMO2 as indicated. In (D), the amounts of d-IP products were adjusted to yield a similar level of unmodified Atxn1 82Q (middle panel).
(F) Levels of Atxn1 82Q-GFP in HeLa cells in the absence or presence of increasing amounts of PML or PML M6.
See also Figure S4.