Supporting Information

Dobler et al. 10.1073/pnas.1202111109

Fig. S1. Survival of HeLa cells (mean of three replicates ± SD) transfected with plasmids bearing the Drosophila melanogaster Na,K-ATPase α-subunit, either as wild-type or with different amino acid substitutions as indicated. Cells were selected over 48 h with 0.1 μM ouabain added to the medium. Bars marked with different letters differ significantly at P < 0.05 (Tukey-Kramer tests).

Table S1. Collecting sites and local hosts of used in this study Species Stage Local food plant Collecting site

Coleoptera Tetraopes tetrophthalmus (Cerambycidae) Adults syriaca Ithaca, NY Alticinae sp. (Chrysomelidae) Adults Asclepias linaria Arizpe, Sonora, Mexico Chrysochus auratus (Chrysomelidae) Adults cannabinum Boulder, CO Chrysochus asclepiadeus (Chrysomelidae) Adults Vincetoxicum officinale Cevio, Ticino, Switzerland Labidomera clivicollis (Chrysomelidae) Adults A. syriaca Ithaca, NY Gastrophysa viridula (Chrysomelidae) Adults Rumex obtusifolius Bremen, Germany Rhyssomatus lineaticollis (Curculionidae) Larvae A. syriaca Toronto, Ontario, Canada Diptera Liriomyzas asclepiadis (Agromyzidae) Larvae A. syriaca Ithaca, NY Liriomyza sp.(Agromyzidae) A. angustifolia Arizpe, Sonora, Mexico Heteroptera Largus sp. (Largidae) Larvae A. angustifolia Arizpe, Sonora, Mexico Oncopeltus fasciatus (Lygaeidae) Adults A. syriaca Ithaca, NY Lygaeus kalmii (Lygaeidae) Adults A. syriaca Ithaca, NY Pyrrhocoris apterus (Pyrrhocoridae) Adults Tiliaceae, Malvaceae Bretten, Baden-Württemberg, Germany oregonensis (Arctiidae) Adults A. cannabinum Boulder, CO Cycnia tenera (Arctiidae) Larvae A. syriaca Ithaca, NY Euchaetes egle (Arctiidae) Larvae A. syriaca Ithaca, NY Lerina incarnata (Arctiidae) Larvae A. linaria Tucson, AZ Pygoctenucha terminalis (Arctiidae) Adults A. cannabinum Boulder, CO futilalis () Larvae A. cannabinum Moab, UT Danaus plexippus (Nymphalidae) Larvae A. syriaca Ithaca, NY Danaus gilippus (Nymphalidae) Larvae A. curassavica Tucson, AZ

Dobler et al. www.pnas.org/cgi/content/short/1202111109 1of2 Table S2. Primers used to amplify the Na,K-ATPase Primer name Ref. Sequence

fATPa 1 5′-ATGACNGTNGCNCATATGTGGT-3′ fATPb 1 5′-GTNATGAAGGGNGCNCCNGA-3′ fATPc 1 5′-ATGGTNACNGGNGATCATCCNAT-3′ rATPa* 1 5′-ATNGGGTGGTCNCCNGTNACCAT-3′ rATPc* 1 5′-ATNGCNGGNACCATGTCNGTNCC-3′ rev-fATPb* 5′-TCNGGNGCNCCCTTCATNAC-3′ sodpump A1368* 5′-TGRTCYTCNGTNGTRTCNGCYTC-3′ † sodpump A903*, 25′-CCYTCNACNGCRTTNRTNGARAA-3′ sodpump B’’ 5′-TNTGGATHGGNGMNATHYTNTG-3′ sodpump C’*5′-ACCATGTTYTTGAASGATTCCATG-3′ sodpump S1957 5′-ATGTCNATGATTGAYCCNCC-3′ sodpump S409 5′-GARCGNGAYGGNCCNAAYGC-3′ sodpump S458 2 5′-GARTGGGTNAARTTYTGYAARAAY-3′ sodpump S475 5′-AARAAYCTNTTYGGGGGTTTYGC-3′ sodpump S772 5′-GTKAARTTYGGYGACCGNATYCC-3′ sodpump S928 5′-TTYTCNAYNAAYGCNGTNGARGG-3′

Numbers refer to the position of the 3′ end in the D. melanogaster gene. *Reverse primer. † Erroneously referred to as A906 in ref. 2.

1. Anderson FE, Córdoba AJ, Thollesson M (2004) Bilaterian phylogeny based on analyses of a region of the sodium-potassium ATPase alpha-subunit gene. J Mol Evol 58:252–268. 2. Labeyrie E, Dobler S (2004) Molecular adaptation of Chrysochus leaf beetles to toxic compounds in their food plants. Mol Biol Evol 21:218–221.

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