IndraLab

Statements

YOD1 binds Flag. 18 / 18
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"In order to exclude the impact of plasmid toxicity on cell growth, we established cell lines expressing doxycycline-induced Flag-YOD1 (Fig. xref )."
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"In a reciprocal immunoprecipitation experiment, overexpressed HA-tagged PML/RAR α could also be detected in the immunocomplex of YOD1-Flag ( xref B)."
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"To determine the substrate proteins regulated by YOD1 in microglia, we transfected NIH/3T3 cells with empty Flag vector or Flag-YOD1 vector and performed immunoprecipitation assay combined with liquid chromatography-tandem mass spectrometry (LC–MS/MS) method to analyze potential substrate proteins of YOD1 ( xref A)."
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"Considering that YOD1 is a deubiquitinating enzyme, we transfected a Flag-YOD1 overexpression plasmid into 786-O cells."
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"We further co-transfected U2OS cells with Flag-YOD1 and Myc-p53 and performed immunoprecipitation using an anti-Myc tag antibody."
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"As expected, we detected clear binding of Myc-p53 to Flag-YOD1 (Fig. xref )."
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"To clarify whether YOD1 can regulate the ubiquitination of PKM2, we co‐transfected HA‐Ub, His‐PKM2 and Flag‐YOD1 or Flag‐CN overexpression plasmids into NIH3T3 cell lines or PC12 cells, and used His magnetic beads to perform Co‐IP experiments, and western blot to detect the ubiquitination of PKM2."
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"As shown in Fig.  xref , Flag-YOD1 and HA-NCOA4 were transfected into HPAEpiC cells as indicated; YOD1 and NCOA4 directly bind to each other under LPS stimulation."
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"To do this, we co-transfected HA-Ub, His-MYH9, and Flag-YOD1 plasmids in NIH/3T3 cells respectively."
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"We co-transfected His-MYH9, Flag-YOD1 and mutant ubiquitin plasmids in NIH/3T3 cells respectively, retaining only the K48 and K63 active sites."
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"Through immunoprecipitation, we found that Flag-YOD1 could bind △C and △M but not DBD or △N mutants (Fig. xref )."
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"Results reveal that exogenously expressed YOD1-Flag and PML/RAR α -HA (both the L and S isoforms) were robustly coprecipitated ( xref A)."
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"For this purpose, U2OS cells were transfected with the control empty vector or plasmid carrying Flag-YOD1, and cell extracts were immunoprecipitated using an anti-FLAG tag antibody."
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"To determine if this was the case, U2OS cells were co-transfected with plasmids carrying HA-Ub, Myc-p53, and Flag-YOD1 and then treated with the proteasome inhibitor MG132."
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"Subsequently, full-length Flag-YOD1 and its various truncations, alone or combined with full-length Myc-ZNF24, were cotransfected into HEK293 cells."
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"Furthermore, we expressed full-length Myc-ZNF24 and its truncations in HEK293 cells, either alone or in combination with full-length Flag-YOD1."
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"Immunoblotting analysis with an anti-p53 antibody revealed the presence of p53 in the immunoprecipitates obtained from cells transfected with Flag-YOD1, but not the empty vector (Fig. xref )."
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"By co‐expressing Flag‐YOD1, His‐PKM2 and mutant ubiquitin constructs (retaining only K48 or K63 lysine residues) in NIH‐3T3 cells, we observed that the HA‐UbK63 plasmid specifically reduced PKM2 ubiquitination (Figure  xref )."
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