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USP13 binds Flag. 41 / 41
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"Co-transfection of NIH/3T3 cells with Flag–USP13, HA–Ub/K63, and His–STING showed that USP13 removed the ubiquitin chain of STING, especially the K63-linked ubiquitin chain ( xref A)."
40487656
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"Next, we generated Flag-USP13 Mut, the USP13 enzymatically inactive triple mutant containing C345A, M664E, and M739E, to observe the restoration of ubiquitination."
36585612
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"HEK293T cells were co-transfected with GFP-tagged HMGB1 (GFP-HMGB1), Flag-USP13, or empty plasmids."
36585612
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"To determine whether USP13 influences the stability of HMGB1, we overexpressed Myc-HMGB1 in HEK293T cells and then increased the number of Flag-USP13 plasmids."
36585612
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"Flag-USP13 Mut, however, failed to stabilize HMGB1 protein levels (Fig.  xref C), indicating that USP13 regulates HMGB1 protein stability."
36585612
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"Next, we transfected Myc-tagged WWP1 (Myc-WWP1), along with Flag-tagged USP13 (Flag-USP13) or an empty vector, into HCC and HEK293T cells, followed by a Co-IP assay to validate the binding between USP13 and WWP1."
40319251
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"Co-IP results further corroborated the exogenous interaction of USP13 and STING in NIH/3T3 cells co-transfected with Flag–USP13 and His–STING ( xref D)."
40487656
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"To determine the ubiquitin linkage patterns of WWP1 by USP13, we co-transfected Myc-WWP1 with HA-Ub mutants with all lysine mutations (K0), or all lysine mutations but one (K6, K11, K27, K29, K33, K48, or K63), along with or without Flag-USP13, into HEK293T cells (Fig.  xref H)."
40319251
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"To further verify the involvement of K29- and K48-linked polyubiquitination of WWP1 by USP13, we co-transfected Myc-WWP1 with wild-type (WT) HA-Ub or HA-Ub with a single lysine mutation (K6R, K11R, K27R, K29R, K33R, K48R, or K63R), along with or without Flag-USP13, into HEK293T cells (Fig.  xref I)."
40319251
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"To further confirm the role of USP13 on METTL3 ubiquitination, HEK 293T cells were co-transfected with Flag-USP13 (either WT or C345A mutant), Myc-METTL3 and HA-Ub."
37151889
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"To test the above hypothesis, several angiogenesis-related cytokines were analyzed for their mRNA levels in CRC cells with sh-USP13 or Flag-USP13 expression ( xref , xref ) and the mRNA levels of VEGFA, VEGFB, VEGFC, VEGFD, PDGFB, PDGFC, THBS1, WNT7B, PGF, bFGF, MMP2, and CXCL9 were quantified."
40746889
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"The protein level of STAT1 was increased in HL-1 cells (Fig.  xref ) and NRCMs (Supplementary Fig.  xref ) overexpressing Flag-USP13, accompanied with the increased P-STAT1 level."
40593642
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"We transfected human NCM460 and murine MODE‐K cells with Flag‐USP13 plasmids, followed by coimmunoprecipitation combined with LC‐MS/MS analysis to account for species differences in protein interactions (workflow shown in Figure xref )."
40679368
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"To ascertain the deubiquitination types of cyclin D1 by USP13, we co-transfected Myc-cyclin D1 and HA-Ub K48 or K63 mutants, in which all lysine residues except the one at position 48 or 63 are replaced with arginine, together with or without Flag-USP13 into HEK293T cells."
37311811
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"To further confirm the role of USP13 in the regulation on IκBα ubiquitination, Flag-USP13 (either WT or C345A mutant), Myc-IκBα as well as HA-Ub were co-transfected in HEK 293T cells."
36915206
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"HL-1 cells and NRCMs were transfected with Flag-USP13 plasmids, respectively, followed by coimmunoprecipitation combined with LC-MS/MS analysis (interactome workflow shown in Fig.  xref )."
40593642
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"In contrast, overexpression of Flag-USP13 decreased the ubiquitination level of cyclin D1 in HEK293T cells and GC cells (Fig. xref )."
37311811
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"We subsequently analyzed VEGFA protein expression in CRC cells expressing sh-USP13 or Flag-USP13."
40746889
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"The above findings demonstrate that CM supplemented with rVEGFA effectively counteracts the inhibitory effect of CM from Flag-USP13 CRC cells against HUVEC angiogenesis."
40746889
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"To test this, we co-transfected 293T cells with Myc-TAK1, Flag-USP13, and HA-Ub."
39033101
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"To explore USP13-mediated degradation of STING protein, HL-1 cells expressing Flag–USP13 were subjected to proteasome inhibitors or autophagy inhibitors."
40487656
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"We transfected Myc-tagged cyclin D1 (Myc-cyclin D1), together with or without Flag-tagged USP13 (Flag-USP13) into HEK293T or different GC cells and carried out exogenous Co-IP assay."
37311811
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"Also, the interaction between exogenous USP13 and GRP78 was confirmed in NIH/3T3 cells co‐transfected with Flag‐USP13 and His‐GRP78 plasmids (Figure  xref ; Figure xref , Supporting Information) in a dose‐dependent manner (Figure  xref )."
40679368
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"We transfected Myc-tagged WT-cyclinD1 or different mutants into HEK293T cells together with HA-Ub and Flag-USP13."
37311811
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"Moreover, in sh-USP13 and Flag-USP13 CRC cells, a positive correlation was observed between USP13 and PTEN expression levels, with AKT phosphorylation levels corresponding to VEGFA expression."
40746889
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"To confirm whether USP13 binds to HMGB1, both Myc-HMGB1 and Flag-tagged USP13 (Flag-USP13) or USP10 and USP15 plasmids were overexpressed in HEK293T cells for co-immunoprecipitation."
36585612
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"Western blot assay confirmed the upregulation of Flag-USP13 expression in heart tissues from mice since TAC 4-week (Supplementary Fig.  xref )."
40593642
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"We examined the overexpression efficiency of AAV9s and showed increased protein levels of Flag-USP13 (Supplementary Fig.  xref ) and Flag-STAT1 (Supplementary Fig.  xref ) in heart tissues."
40593642
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"HEK293T cells were co-transfected with both Myc-HMGB1 and Flag-USP13 plasmids along with the HA-Ub plasmid for 48 h and then treated with MG132, a proteasome inhibitor for the next 18 h before harvest."
36585612
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"As shown in Fig. xref , Myc-cyclin D1 could be detected in Flag-USP13 immunoprecipitates using an anti- FLAG antibody in the cells transfected with Myc-cyclin D1 and Flag-USP13."
37311811
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"Subsequently, to further explore the interaction between USP13 and ALDOA, we transfected plasmids encoding Flag-USP13 and Ha-ALDOA into 293T cells and found the interaction between USP13 and ALDOA with co-immunoprecipitation approach ( xref F and G)."
41496210
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"As USP13 is a DUB that stabilizes ALDOA via proteasome-mediated degradation, we next investigated whether USP13 participates in the ubiquitination of ALDOA in 293T cells cotransfected with Ha-ALDOA, Myc-Ub and Flag-USP13."
41496210
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"HEK293T cells were co-transfected with Myc-tagged HMGB1, HA-Ub, Flag-USP13, or Flag-USP13 Mut and treated with MG132."
36585612
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"Flag-USP13 was also detected in the Myc-WWP1-binding complex with the Myc antibody (Fig.  xref D)."
40319251
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"The levels of HMGB1 K48-linked deubiquitination induced by Flag-USP13 overexpression were significantly recovered by an inactive mutant of Flag-USP13 Mut (Fig.  xref C)."
36585612
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"Consistently, Flag-USP13 could be pulled down by Myc-cyclin D1 with an anti-Myc antibody (Fig. xref )."
37311811
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"We cotransfected 293T cells with Ha-ALDOA and various concentrations of Flag-USP13."
41496210
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"We reconfirmed that K48-linked HMGB1 ubiquitination was reduced by USP13 overexpression when HEK293T cells were cotransfected with Myc-HMGB1 and HA-Ub K48 plasmids in the presence or absence of Flag-USP13 (Fig.  xref F)."
36585612
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"Expression plasmids for Flag‐USP13 (human or mouse), USP13‐C343A (mouse), His‐GRP78 (mouse), His‐GRP78‐K327R (mouse), HA‐Ub (mouse), HA‐K63 (mouse), and HA‐K48 (mouse) were constructed by Genechem (Shanghai, China)."
40679368
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"NCM460 or MODE‐K cells were transfected with Flag‐empty vector (EV) or Flag‐USP13 (USP13 OE ) plasmids, followed by stimulation with LPS (5 µg mL −1 )."
40679368
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"The exogenous interaction between USP13 and STAT1 was examined in NIH/3T3 co-transfecting Flag-USP13 and STAT1 plasmids (Fig.  xref )."
40593642