Epidermal and Intestinal PGP-9 Expression

Plasmids for transgenesis were assembled using the NEB HIFI DNA Assembly Kit (New England Biolabs Inc., Ipswich, MA, USA) according to the manufacturer’s instructions into the pUC19 vector linearised with SmaI (ThermoFisher, Waltham, MA, USA). Plasmid constructs were Cel-col-19p::Pun-pgp-9::FLAG::Cel-unc-54_3′-UTR utilising the col-19 promotor [39 (link)] to drive epidermal Pun-PGP-9 expression (Supplementary Figure S3a) and Cel-ges-1p::Pun-pgp-9::FLAG::Cel-unc-54_3′-UTR (Supplementary Figure S3b) utilising the ges-1 promotor [38 (link)] to drive intestine-specific Pun-PGP-9 expression. The C. elegans unc-54 3′-UTR [41 (link)] and the Pun-pgp-9 cDNA [23 (link)] were amplified from verified plasmids, while the 3′ end primer for the Pun-pgp-9 amplification introduced an in-frame FLAG-tag (DYKDDDDK) before the stop codon (all primers in Supplementary Table S3). The C. elegans promotors col-19p and ges-1p [38 (link)] were amplified from genomic DNA extracted from the Bristol N2 strain [39 (link)]. A co-injection marker plasmid (pPD118.33) driving pharyngeal GFP expression was used (Addgene L3790 plasmid #1596 was a gift from A. Fire). Sequences of all constructs were confirmed by Sanger-sequencing (LGC Genomics, Hoddesdon, UK).