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This file contains a full description of the statistical analyses, as well as their out-put. A supplementary figure about the effect of diet on pupal weight and body size of the samples is also provided.

Additional file 1: Table S1. Clone and primer list. Table S2. Yeast expression constructs. Table S3. Assigned 1C and 13C NMR peaks for compound 5 and 6. Figure S1. Comparison of the mass spectra of TMS derivatized TwOSC products from tobacco leaf co-infiltration experiments, with TMS derivatized authentic standards. Figure S2. CYP712K1 and CYP712K2 oxygenate the triterpene products of TwOSC2 and TwOSC3 in tobacco. Two leaf discs (Ø3 cm) of leaves co-expressing the indicated enzymes were extracted in 1 mL ethyl acetate. A 0.15 mL aliquot of this extract was dried, TMS derivatized and analyzed by GC–MS. None of the novel peaks 7-9 had mass spectra or retention times matching any of the analyzed authentic standards (betulinic acid, hederagenin, or oleanolic acid; not shown). Figure S3. Comparison of LC–ESI–MS and LC-APCI-MS (base peak chromatograms) for the reliable quantification of friedelin and polpunonic acid. Authentic standards of friedelin and polpunonic acid (30 µg/mL) were analyzed using the same LC–MS equipment and program with exception of ionization probe (ESI; black trace and APCI; red trace).

Western blot methods

Primer sequences, restriction sites are underlined, and 6xHIS-tag is in italics