Fig. 2

Schematic of the workflow. a The GTA tag is added to a gene of interest using recombineering. b Transgenic C. elegans strains expressing the GTA-tagged protein are generated by injection followed by gamma irradiation-mediated integration of the extrachromosomal array. c Transgenic strains are crossed with strains expressing BirA from a tissue-specific promoter, and with a genetic null mutant if appropriate. d The transgenic strains are grown in triplicate in liquid culture. e Affinity purification is performed on whole-animal lysates. The bait protein with any interacting proteins is subsequently cleaved off the beads by Tobacco etch virus (TEV) protease. f The samples are analyzed by tandem mass spectrometry (MS/MS) to identify the proteins they contain