Fig. 3: CAND2 promotes the exchange of F-box proteins (FBPs) but less efficiently than CAND1 does.

A, B CAND2 promotes the assembly of new SCF complexes by accelerating the disassembly of pre-existing SCFs. A Schematic diagram of the experimental design. DKO cells expressing endogenous ^3xFLAGCUL1 were subjected to IP with an anti-FLAG antibody in the presence of excess SKP1^Sortase·SKP2^ΔLRR chase protein and equal concentrations of recombinant CAND1 or CAND2. Samples were immunoblotted with the indicated antibodies (B). Representative results from three independent experiments are shown. C SILAC-based quantitative proteomic assay assessing FBP exchange levels in cell lysates. The experimental procedure is illustrated (upper flowchart), and general profiles of FBP exchange under different conditions are shown (bottom graph). Black bars: median exchange. D Exchange levels of CSN and the common subunits of SCF. Results from three biological replicates are shown. H/L ratio: heavy/light ratio. E Exchange levels of FBPs detected in all four experimental groups. Results from three biological replicates are shown. F Exchange levels of FBPs that displayed increased exchange only in response to CAND1. Data are presented as mean ± SEM; n = 3 biological replicates except for FBXO5 in the 1x CAND1 group (n = 2 biological replicates). G Exchange levels of FBPs detected only in the absence of CAND1. Results from three biological replicates are shown. Source data are provided as a Source Data file.