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OTUD3 binds FlaG. 10 / 10
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"In addition, OTUD3 C76A catalytic mutation (as FLAG-OTUD3 CA) does not obviously affect the interaction between HA-cGAS and FLAG-OTUD3 ( Figure S3 C)."
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"Moreover, under HT-DNA activation, association of endogenous cGAS with FLAG-OTUD3 or endogenous OTUD3 was enhanced, probably due to the higher expression of endogenous cGAS induced by DNA activation ([MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"By using 90-bp dsDNA in DNA shift assay ( Figures 6 C and 6D), we further established that the Kd (equilibrium dissociation constant) of 90-bp dsDNA binding to OTUD3 is about 0.5 μM. To examine whethe[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"19 According to the study showing that OTUD3 associates with mitochondrial antiviral-signaling protein (MAVS) to suppress anti-RNA viral immune response, 17 we also performed coIP experiments using F[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"29 , 30 , 31 To examine whether OTUD3 affects cGAS enzymatic activity, FLAG-cGAS was purified from 293T shOTUD3#1cells and treated with FLAG-OTUD3 WT or CA in vitro , followed by cGAS enzymatic assay."
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"From in vitro assays using FLAG-OTUD3 WT/CA purified from 293T cells and FLAG-cGAS bearing polyUb purified from 293T shOTUD3#1 cells (either in native condition or denaturing condition), it is clear t[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"As shown in Figure 1 I, FLAG-OTUD3 and GFP-cGAS are colocalized in cytosolic GFP-cGAS-DNA complex under DNA activation in HeLa cells."
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"Furthermore, in THP-1 FLAG-OTUD3 cells, besides some low overlap of FLAG-OTUD3 and endogenous cGAS in resting conditions, concentrated FLAG-OTUD3 resides in endogenous cGAS-DNA complex not only under [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"As shown in Figure 6 E, FLAG-OTUD3 indeed pulled down 500-bp DNA the same as FLAG-cGAS, while GFP-FLAG could not."
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"To investigate whether OTUD3 interacts with cGAS, we carried out coIP from cells using FLAG-OTUD3 together with hemagglutinin (HA)-cGAS, and found HA-cGAS in FLAG-OTUD3 IP sample but not in control im[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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