Fig. 1
From: EGFR feedback-inhibition by Ran-binding protein 6 is disrupted in cancer
Importin β-like protein RanBP6 interacts with EGFR. a Left panel, schematic representation of EGFR immunoaffinity purification and LC-MS/MS analysis in A431. Right panel, plot showing the ten top categories of the gene ontology enrichment analysis of the EGFR-associated proteins. b List of the unique peptides for RanBP6 identified in the MS analysis and the replicate in which the peptide was identified are indicated in the table. c Co-immunoprecipitation of EGFR and V5 epitope-tagged RanBP6-V5 in A431 cells. Top panel, IP using V5 antibody; bottom panel, immunoblot of whole-cell lysates (WCL). d Conserved domains within the family of importin β-related proteins. RanBP6 includes an importin β-like N-terminal domain (Imp. N-ter), seven HEAT repeats, and a putative Ran-binding domain (RBD). The number to the right of each protein shows the total number of amino acids. e RanBP6 interacts with nuclear but not cytoplasmic Ran-GTPase. Subcellular fractionation of HEK-293T cells (right panel) shows that Ran is present in both nuclear and cytoplasmic compartments, but only interacts with RanBP6 in the nuclear fraction (left panel). f Venn diagram representing overlapping proteins between the RanBP6-V5 and the EGFR immunoaffinity purifications. See Supplementary Data 5. g GST pulldown assay confirms the interaction of RanBP6 with importin-α1, importin-β1, RanGAP1, and nuclear pore complex 93 (NUP93) in HEK-293T whole-cell lysates
