IndraLab

Statements

USP48 binds Flag. 5 / 5
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"To assess the dose-dependent effect of USP48 on HEG1 deubiquitination, we overexpressed increasing amounts of Flag-USP48 in 293 T cells co-transfected with HA-HEG1, His-Ub, and Flag-vector control."
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"Finally, to determine the specific type of ubiquitin linkage removed by USP48, we transfected 293 T cells with HA-HEG1, Flag-USP48, and various His-tagged ubiquitin constructs: wild-type ubiquitin (Ub-WT), K48-only (Ub-K48), and K63-only (Ub-K63)."
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"To explore whether USP48 regulates HEG1 expression, we transfected 293 T cells with increasing amounts of Flag-USP48 plasmid and harvested proteins 48 hs post-transfection."
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"293 T cells were transfected with Flag-USP48 or Flag-USP48-C98S along with His-Ub, and ubiquitinated HEG1 levels were examined following Co-IP."
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"Cells were co-transfected with HA-tagged HEG1, His-tagged ubiquitin (His-Ub), and either wild-type Flag-USP48 or the catalytically inactive mutant Flag-USP48-C98S. Forty-eight hours post-transfection, and following 8-hour MG132 treatment to inhibit proteasomal degradation, cell lysates were subjected to Co-IP using anti-HA antibodies."
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