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OTUD5 binds FlaG. 8 / 8
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"To test whether OTUD5 also interacts with SPT16 in an unbiased method, we performed FLAG-OTUD5 IP and GST-OTUD5 pulldown experiments from HeLa S3 cells and subjected the eluate to mass spec analysis (Figure xref )."
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"At last, the PLA assay showed that FLAG-OTUD5 WT interacted with endogenous UBR5, measured by increased PLA spots, but the FLAG-OTUD5 ΔN17 mutant did not (Figure xref ), further supporting the conclusions."
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"First, we found that expressing the FLAG-OTUD5 WT reversed (repressed) the YFP-MS2 reporter expression at the Fok1-induced DSB sites in OTUD5 knockdown cells (Figure xref )."
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"To identify OTUD5 target(s) during NF-κB activation, we analyzed FLAG–OTUD5 associated protein identified by LC-MS."
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"However, neither ΔN17 nor ΔUIM mutant were able to rescue the phenotype (Figure xref ; presence of FLAG-OTUD5 mutants at the spots indicate the normal expression and localization)."
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"Below panel shows the expression confirmation of the FLAG-OTUD5 truncates."
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"The depleted UBR5 proteins were effectively rescued by expressing FLAG-UBR5 wild-type (WT), but not by a catalytically inactive FLAG-OTUD5 mutant (C224S) ( xref ) (Figure xref ), suggesting that the deubiquitinating activity of OTUD5 is necessary for preserving the UBR5 stability."
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"Indeed, we have observed this in several ways; foci of endogenous OTUD5 and GFP-OTUD5 were detected at UVC-induced γH2AX spots (Figure xref and  xref , respectively; FLAG-OTUD5 in xref )."
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