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USP22 binds FlaG. 16 / 16
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"Consistent with our LacO array recruitment experiment, ( xref ) mCherry-PALB2 WD40 4X could not pull-down FLAG-USP22 WT while mCherry-PALB2 WD40 WT robustly bound FLAG-USP22 WT ."
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"The fragment mCherry-PALB2 Nterm that does not contain the WD40 domain was not able to pull-down FLAG-USP22 WT validating our previous LacO array data ( xref )."
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"Subsequently, after transfecting the cells with the USP22-FLAG and USP22–C185S-FLAG plasmids under the NG and HG conditions, we detected the ubiquitination level of Snail1."
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"To determine whether USP22 could affect the development of DKD, we used a USP22 overexpression plasmid (USP22-FLAG) and a USP22 deubiquitinase activity mutant plasmid (USP22–C185S-FLAG) to transfect t[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Moreover, the results of immunofluorescence experiments showed that transfected FLAG-USP22 (FITC, green) and endogenous ZEB1(Cy3, red) were distributed in the nucleus in HCC-derived cells or HEK293 cells (Fig. xref and Supplementary Fig. S xref D)."
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"WB results showed that USP22 expression significantly increased in NRK-52E cells transfected with USP22-FLAG and USP22–C185S-FLAG ( Fig. 3 A), which verified the success of USP22 overexpression."
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"Therefore, we detected the ubiquitination level of Sirt1 to verify the enzyme activity of USP22 plasmids in NRK-52E cells transfected with USP22-FLAG and USP22–C185S-FLAG."
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"As shown in the data in Fig. 3 B, USP22-FLAG rather than USP22–C185S-FLAG could reduce the Sirt1 ubiquitination level, indicating that the USP22 plasmids functioned normally in the EMT model."
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"Furthermore, overexpression of siRNA resistant FLAG-USP22 in U2OS FOKI cells rescues the localization of BRCA2, PALB2, and Rad51 to the array after DNA damage."
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"In HG-treated NRK-52E cells, USP22-FLAG overexpression further increased USP22 expression ( Fig. 3 C), downregulated E-cadherin and ZO-1expression, and upregulated Vimentin expression ( Fig. 3 D)."
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"Cell-scratch test results showed that in HG-treated NRK-52E cells, USP22-FLAG overexpression significantly increased cell migration, whereas USP22–C185S-FLAG overexpression had no such function ( Fig.[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"In NRK-52E cells transfected with USP22-FLAG and USP22–C185S-FLAG, WB results showed that USP22-FLAG overexpression, not USP22–C185S-FLAG overexpression, further promoted HG-induced Snail1 upregulatio[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"USP22-FLAG plasmid, USP22–C185S-FLAG plasmid ( Kobayashi et al., 2015 ), and USP22-shRNA plasmid were all purchased from Genechem (Shanghai, China)."
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"The wild-type (WT) USP22-FLAG plasmid and the deubiquitinase activity mutant USP22–C185S-FLAG plasmid were constructed with the assistance of Genechem (Shanghai, China)."
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"Using an mCherry tagged version of this mutant (mCherry-PALB2 WD40 4X ) we performed an IP experiment in H1299 lung adenocarcinoma cells to assess its ability to pull down FLAG-USP22 WT ( xref )."
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"The overexpression of USP22-FLAG, rather than USP22–C185S-FLAG, further reduced the ubiquitination level of Snail1, which exerted a superimposing effect with HG ( Fig. 7 D)."
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