Fig. 1: PUP-IT identifies protein interactors of RACK1 and TPLATE in plant cells.

a Schematic of the PUP-IT proximity labeling approach [Created in BioRender. Persson, S. (2025) https://BioRender.com/z33f531]. FLAG-Pup(E) is attached to Prey protein mediated by PafA fused to a bait protein. b Vectors designed to express the FLAG-Pup(E) and PafA fused to GFP/AtRACK1a/NbRACK1, with cassettes for PafA and Pup(E) expressions present in one binary vector. c Venn diagram generated from enriched proteins of transiently expressed AtRACK1a and NbRACK1 as PafA-baits in N. benthamiana compared to PafA-GFP, respectively. d Molecular function enrichment of proteins in the overlap of (c). e A PPI network of putative RACK1 interactors that is also supported by interactions from the String database (https://string-db.org/). Nodes indicate proteins and edges indicate interactions, with line thickness indicating the strength of data support. f Vectors designed to express STREPII-FLAG-Pup(E) under a Dexamethasone (DEX)-inducible promoter, combined with constitutive expression of PafA fused to TPLATE. g iBAQ, a measure for the abundance of TPLATE subunits. iBAQ of TPLATE subunits were normalized to the bait, TPLATE, for the different experiments (as indicated below).