IndraLab

Statements

OTUD4 binds Flag. 8 / 8
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"Furthermore, co-transfection of His-ZMYND8, Flag-OTUD4, and HA-Ub plasmids in HEK-293T cells demonstrated that overexpression of OTUD4 markedly reduced ZMYND8 ubiquitination ( xref B), confirming that OTUD4 promotes ZMYND8 stability through a deubiquitination mechanism."
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"To investigate underlying mechanisms by which OTUD4 promotes the migration and invasiveness of TNBC cells, MDA-MB-231 cells stably expressing Flag-OTUD4 were used to perform tandem affinity purificati[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"OTUD4 was over-expressed by transfection with Flag-OTUD4 plasmid."
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"These mutants were co-transfected with Flag-OTUD4 and HA-Ub into HEK-293T cells, followed by co-immunoprecipitation (Co-IP) assays."
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"Experiments with MiaPaCa-2 and Panc01 cells provided definitive proof of complex formation between Flag-OTUD4 and HA-RPL6 upon their simultaneous expression (Fig.  xref A)."
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"Moreover, when co-transfected with Flag-OTUD4, overexpression of OTUD4 decreased ubiquitination of ZMYND8 wild-type (WT; human), K356R (human), K508R (human), WT (mouse), K380R (mouse), and K532R (mouse) mutants, but failed to reduce ubiquitination of the ZMYND8 K294R (human) and K318R (mouse) mutants ( xref B)."
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"It is worth noting that the slightly elevated His-ZMYND8 levels observed in the Flag-OTUD4 input lanes reflect the stabilizing effect of OTUD4 on ZMYND8, consistent with our findings in xref F–G, and do not influence the interpretation of ubiquitination levels obtained from the His immunoprecipitation."
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"To verify an interaction between OTUD4 and SHBs, Huh7 cells were co-transfected with plasmids encoding SHBs-Strep-Flag and OTUD4-Flag."
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