Figure 4.

Pull-down assays using NZF variants containing point mutations in the Φ position of the TF/Φ motif to assess their mono- or diubiquitin binding. (A) Pull-down assays using the GST-fused TAB2-, Npl4- and Vps36-NZF mutants to assess their monoubiquitin binding. The bound monoubiquitin molecules were analysed by tricine SDS–PAGE and stained with Coomassie Brilliant Blue. (B) Pull-down assays using GST-fused TAB2-, Npl4- and Vps36-NZF mutants to assess their binding with K63-Ub₂, K48-Ub₂ or linear Ub₂. The bound diubiquitin molecules were analysed by SDS–PAGE and stained with Coomassie Brilliant Blue.