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UCHL3 binds Flag. 10 / 10
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"Consistently, we co-overexpressed Flag-UCHL3, Myc-NRF2, and HA-ubiquitin (HA-Ub) in HEK293 and the ubiquitination assay also demonstrated that UCHL3 could stabilize NRF2 protein via deubiquitination ([MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"To investigate the interaction between Uch-L3 and Smad1, HA-Smad1 and Flag-Uch-L3 plasmids were transfected into HEK 293 cells."
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"Immunoblotting of cell lysates showed that UCHL3 coimmunoprecipitated with CTNNB1 when Flag-UCHL3 and HA-CTNNB1 were coexpressed in 293 T cells (Fig.  xref A)."
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"Uch-L3 protein (Flag-Uch-L3) was detected in the eluent of the immuno-complex by western blot using anti-Flag antibody ( Fig. 1 A)."
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"1 Experimental Models: Cell Lines HEK293T Laboratory of Michiel Vermeulen N/A HeLa Cells Spruijt et al., 2013 N/A IB10 Cells Spruijt et al., 2013 N/A NPCs Spruijt et al., 2013 N/A HEK293T-EGFP-TAB-NZF[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Reciprocally, the co-IP against Flag-Uch-L3 demonstrated that the eluent contained HA-Smad1 ( Fig. 1 B)."
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"As shown in Fig. 1 D, the endogenous Smad1 was also detected in the immunoprecipitated complex of Uch-L3 (Flag-Uch-L3)."
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"We first cotransfected different amounts of Flag-UCHL3 (0, 200, 400, 600 and 800 ng) into 293 T cells with HA-CTNNB1."
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"Then, we confirmed these interactions by co-IP assay and GST pull-down assay and found that inactive UCHL3 (Flag-UCHL3 S75A) can bind POLD4 directly, indicating that the enzyme activity of UCHL3 was not crucial for their interaction (Fig.  xref D, E)."
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"For Co-IP and ubiquitination assay in HEK293T cells, HEK293T cells were co-transfected with Myc-NRF2, HA-Ub, and Flag-UCHL3."
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