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. 2013 Dec 19;289(6):3444–3456. doi: 10.1074/jbc.M113.507541

FIGURE 2.

FIGURE 2.

Stimulation of USP-46 catalytic activity by WDR-20 and WDR-48 in vitro. Representative immunoblot analysis showing the effect of Ub-VME on FLAG·USP-46 complexes isolated from HEK293T cells. A, the wild type (FLAG·USP-46(WT)) was transfected either alone or together with WDR-48 alone, WDR-20 and WDR-48 together, or WDR-20 alone. B, FLAG·USP-46(C>A) (catalytically inactive) was transfected either alone or together with WDR-20 and WDR-48. Wild-type (FLAG·USP-46(WT)) (A) or catalytically inactive (FLAG·USP-46(C>A)) (B) complexes were immunoprecipitated and incubated with or without HA-Ub-VME. After halting the reaction, adduct formation was assayed by immunoblotting against FLAG·USP-46. Similar results were obtained in more than three independent experiments.