IndraLab

Statements

USP28 binds Flag. 18 / 18
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"We used USP28-KO clones exhibiting reduced MAST1 protein level and the cells were reconstituted either with Flag-USP28 or Myc-MAST1 for further functional experiments (Fig.  xref A; Supplementary Fig. 8)."
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"293T cells were co-transfected with Flag-USP28, His-SUMO2, GFP-SENP1 WT or SENP1 catalytic mutant (SENP1 CA)."
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"To this end, we transfected both A549 and HEK293 cells with increasing concentrations of Flag-USP28 and analyzed MAST1 protein level."
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"To examine the deubiquitinated modification of TRIM21 by USP28, we evaluated the ubiquitination level of TRIM21 in HL-1 cells expressing Flag-USP28 (USP28 OE ) and found that USP28 OE promoted the deubiquitination of TRIM21 (Fig. xref D)."
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"However, the half-life of MAST1 protein was rescued when USP28-KO cells were reconstituted with Flag-USP28 (Fig.  xref J, lane 9–12; Supplementary Fig. 4A, lane 9–12), while USP28CA failed to increase the half-life of MAST1 protein (Fig.  xref K, lane 9–12; Supplementary Fig. 4B, lane 9–12)."
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"Here, we performed a transcriptome in Ang II-challenged HL-1 cells expressing Flag-USP28 (USP28 OE ) and showed that USP28 OE negatively regulates antioxidant activity and biological oxidations pathways (Fig. xref A and xref A-B)."
39431010
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"To test this, a Flag-USP28 expression construct was transfected into T24T cells; as expected, USP28 over-expression led to increased CD44 protein expression ( xref )."
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"Flag-USP28 and GFP-SENP1 plasmids were co-transfected into 293T cells for 36 h."
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"Flag-USP28 WT or Flag-USP28 K99R was co-transfected with His-ubiquitin, FBXW7, and HA-HIF-1α into 293T cells for 36 h, and MG132 was added 4 h before cells were harvested."
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"Cells were harvested, and the immunoprecipitation of Flag-USP28 revealed the presence of GFP-SENP1 protein (Fig.  xref a), and SENP1 was also present in immunoprecipitates of endogenous USP28 in HCT116 cell lysate (Fig.  xref b)."
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"The loss of USP28 resulted in lower cell viability compared with the mock control, while reconstitution with Flag-USP28 or Myc-MAST1 increased cell viability (Fig.  xref B; Supplementary Fig. 10)."
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"We found that bacterial purified GST-SENP1 protein was able to capture the Flag-USP28 protein purified from 293T cells."
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"Furthermore, USP28-depleted cells showed an increase in sub-G1 populations whereas reconstitution with Flag-USP28 or Myc-MAST1 decreased sub-G1 populations (Fig.  xref C; Supplementary Fig. 11)."
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"USP28-KO clones from both A549 and H1299 cells showed increased γH2AX foci formation compared with mock control, while reconstitution with Flag-USP28 or MAST1 reduced γH2AX foci formation (Fig.  xref D; Supplementary Fig. 12A)."
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"To investigate the impact of USP28 SUMOylation on HIF-1α, we generated Flag-USP28 K99R mutant plasmid."
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"For protein purification of ubiquitinated Flag-H2AX, Flag-Dub3 WT/CI and Flag-USP28, 293T cells were transfected with the corresponding expression vectors."
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"Furthermore, we observed that the sgRNA-mediated reduced expression of MAST1 protein was rescued upon addition of Flag-USP28 in USP28-depleted cells both at endogenous (Fig.  xref H, lane 4 vs. lane 2) and exogenous levels (Fig.  xref K, lane 5 vs. lane 3)."
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"Co-immunoprecipitation analysis of exogenous Flag-USP28 and Myc-MAST1 in HEK293 cells showed that both proteins interact with each other (Fig.  xref B)."
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