Fig. 4
Identification of a short linear motif (SLiM) in CtNup145C-NTD that recruits the CtNup82 complex. a Schematic representation of ScNup145C and CtNup145C and the design of truncated CtNup145C-A and CtNup145C-B motifs; see also Supplementary Fig. 2. The crosslinking hotspots found in CtNup145C-NTD (Fig. 1b) are indicated by dashed pink lines. DIM domain invasion motif. b In vitro binding assay with Flag-immobilized CtNup145C-A, CtNup145C-B, and CtNup82 complex (Flag-CtNup159C–Nup82–Nsp1C) and soluble GST-CtNup145C-A, GST-CtNup145C-B, CtNup82 complex, and/or CtNup145N-ΔFG. Shown is the analysis of SDS eluates by SDS-PAGE and coomassie staining. The image section marked by the dashed box was subjected to image processing to increase the visibility of the small bait peptides CtNup145C-A-Flag and CtNup145C-B-Flag. c In vitro binding assay with Flag-immobilized CtNsp1C, CtNup159C, CtNup82, and CtNup82 complex (Flag-CtNup159C–Nup82–Nsp1C) and soluble GST-CtNup145C-B or CtNup145C. The experiments were performed at least twice with consistent results. The asterisk indicates bands corresponding to the anti-Flag light chain; M marker, Mock purification buffer with E. coli whole cell lysate, MW molecular weight. Uncropped gel images are shown in Supplementary Fig. 4e, f
