Co-expression of HA-CLC-1 and the Myc vector was used as the control experiment. (B) Co-immunoprecipitation of Myc-CLC-1 and Flag-DDB1 in HEK293T cells. Co-expression of Flag-DDB1 and the Myc vector was used as the control experiment. (C) Lack of co-immunoprecipitation of Myc-CLC...
One of the characteristics of the Nedd8-cullin dimerization mechanism is that the dimeric CRL complex must have equal levels of neddylated and unneddylated cullins. Immunoprecipitation of the CUL3 substrate cyclin E pulls down roughly equivalent levels of neddylated and unneddylated CUL3 [37]. ...
Cullin-associated and neddylation-dissociated protein 1 (CAND1) is a critical regulator of CRLs. It can modulate the formation rate of substrate receptor complex e.g. S-phase kinase-associated protein 1 (Skp1)-Cullin1-F-box protein (SCF), and then control the degradation rate of substrate p...
proteins do not play a role in this function of either biv or mvv vif. co-immunoprecipitation assays further verified the direct interaction of biv vif with bteloc and btcul2, but not with btcbf-β or btcul5. mvv vif also was confirmed to directly interact with oaeloc and oacul5, ...
Immunoprecipitation of endogenous CSN1 from nuclear extract weakly but consistently pulled down endogenous SAP130 (Fig. 1D). In addition, recombinant GST-CSN1, but not GST itself, could pull down in vitro translated SAP130 from the lysate (Fig. 1E). Similarly, another member of DDB1 family,...
Using HIV infectivity, virion incorporation, immunoprecipitation, and in vitro ubiquitin conjugation assays, we show that the ability of Vif to inhibit antiviral activity of APOBEC3 proteins positively correlates with its ability to bind and ubiquitinate these proteins by a Vif–Cullin5–ElonginB–...
(E) Radiolabeled lysates containing Cdc4p (lanes 1–6) were mixed with radiolabeled Skp1pHA (lanes 3–6) or Cdc53p (lanes 5 and 6) lysates prior to addition of MBP-Sic1pMH6, followed by immunoprecipitation using anti-MBP antibodies. To test whether this set of recombinant proteins was ...
Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (P0013B, Beyotime Biotechnology, Shanghai, China) before WB analysis. For co-IP, cells were lysed in mild RIPA lysis buffer (P0013D, Beyotime). The supernatant was first incubated with an antibody overnight, subsequently incubated ...
These increases were confirmed by reciprocal immunoprecipitation/immunoblotting experiments (Fig. 1d; Supplementary Fig. S1c). Conversely, Pof15p levels were 3.5-fold lower in Cul1p complexes isolated from Dknd1 cells (Fig. 1c) and the fractional occupancy for this FBP was reduced from 4.6% to...
To determine how CUL5 regulates IFN-β production via OGT, we first investigated whether CUL5 interacts with OGT. Using co-immunoprecipitation (Co-IP), we found that CUL5 interacts with OGT in HEK293T cells (Fig.6a). CUL5 contains a long stalk-like amino-terminal domain (NTD) comprising...