IndraLab

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OTUD7B binds Cebpb. 5 / 5
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"The luciferase reporter containing wildtype TCF/LEF binding sites (TOPflash) and mutated TCF/LEF binding sites (FOPflash) was transfected with SFB-USP36 or SFB-OTUD7B. Compared to FOPflash luciferase levels, both USP36 and OTUD7B increased the luciferase activities (TOPflash), but OTUD7B significantly enhanced LEF1 activity to a greater extent (6.64-fold) than USP36 did (2.04-fold) ( xref C)."
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"We prepared cDNA samples from HEK293T cells transiently transfected with either SFB-GFP (control) or SFB-OTUD7B, and performed qPCR analysis."
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"We co-transfected these constructs and full-length SFB-OTUD7B into HEK293T cells and pulled down SFB-OTUD7B with S-protein agarose beads."
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"Next, we generated three deletion mutants of OTUD7B: OTUD7BΔZnF (Zinc Finger domain deleted, aa 1-1077), OTUD7BΔOTU (Ovarian Tumor domain deleted, aa 1-564 & 1078-2532), and OTUD7BNt (UBA domain-containing, 1-564) using the SFB-OTUD7B plasmid ( xref C) and attempted to identify the LEF1-binding region on OTUD7B. Similarly, we co-transfected these constructs and full-length MYC-LEF1 into HEK293T cells and pulled down full-length and mutant SFB-OTUD7B with S-protein agarose beads."
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"To investigate this, we co-transfected MYC-LEF1 with SFB-OTUD7B or SFB-GFP plasmids into HEK293T cells and separated the cytoplasmic and nuclear fractions of the cell lysates."
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