Fig. 2: RepID-CRL4 interacts with BUB3 during mitosis. | Nature Communications

Fig. 2: RepID-CRL4 interacts with BUB3 during mitosis.

From: The RepID–CRL4 ubiquitin ligase complex regulates metaphase to anaphase transition via BUB3 degradation

Fig. 2

a BUB3 interacts with RepID. Soluble nuclear and chromatin-bound fractions were extracted from RepID FL-stable U2OS cells and immunoprecipitated using an anti-FLAG antibody, followed by mass spectrometric analysis. b The WD40 ___domain of RepID is required for interaction with BUB3. Top, constructs of RepID variants used in this analysis. Bottom, nuclear and chromatin-bound fractions were extracted from U2OS cells expressing the RepID FL, F2–5, and Δ3 fragments and were immunoprecipitated with FLAG antibody and analyzed by immunoblotting. c BUB3 interacts with RepID-CRL4. Asynchronized HCT116 RepID WT cells were lysed, and BUB3-bound proteins were immunoprecipitated with anti-BUB3 antibody, followed by immunoblot analysis. d BUB3 interacts with RepID-CRL4 during mitosis. K562 RepID WT cells were fractionated by elutriation, and a mixture of soluble nuclear and chromatin-bound fractions were immunoprecipitated with anti-BUB3 antibody, followed by immunoblot analysis.

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