Fig. 2: Functional characterisation of EpSS-L1 and EpSS-L2 in N. benthamiana.

A Four week old N. benthamiana plants infiltrated with: Empty Vector (EV), A. thaliana Squalene Synthase (AtSS, GenBank accession BT003419.1), E. peplus Squalene Synthase-like1 (EpSS-L1, GenBank locus tag M5689_021805), E. peplus Squalene Synthase-like2 (EpSS-L2, GenBank locus tag M5689_021806) with and without truncated A. thaliana HMG-CoA reductase (HMGRt, GenBank accession J04537.1). Infiltrated leaves were extracted and analysed by LC-MS in parallel with E. peplus latex-purified peplusol standard. Extracted Ion Chromatograms (EIC) shown for m/z 427.4 showing peplusol (1) and 2,3-oxidosqualene (3) B Peplusol levels were quantified by LC-MS in N. benthamiana leaves infiltrated with gene combinations as on panel A, against a standard curve generated from purified peplusol. n.d.: not detectable, Error bars: SEM (n = 5). C Squalene levels were quantified by GC-MS in N. benthamiana leaves infiltrated with gene combinations as on panel A, n.d.: not detectable, Error bars: SEM (n = 5, where n is the number of biological replicates). Statistically significant (one-sided t-test) changes between control (EV) and candidate genes indicated by asterisks (*: p-value < 0.05,  **: p-value < 0.01). D, E 2,3-oxidosqualene levels were quantified by LC-MS in N. benthamiana leaves infiltrated with gene combinations as on (A) against a standard curve generated from purified 2,3-oxidosqualene. Error bars: SEM (n = 5, where n is the number of biological replicates). Statistically significant (one-sided t-test) changes between control (EV) and candidate genes indicated by asterisks (**: p-value < 0.01). Source data for (B–E) are provided as a Source Data file.