Tables S1 - S2 and Figure S1 from New ways to acquire resistance: imperfect convergence in insect adaptations to a potent plant toxin

Table S1: RT-qPCR primers designed for P. aegrotus. NKA - Na,K-ATPase, NS - nervous system.; Table S2: Mutagenesis primers to reconstruct the two versions of the P. aegrotus (Paeg) first extracellular loop starting from a D. melanogaster construct. NT nervous system.; Figure S1: Translated amino acid alignment of the Na,K-ATPase alpha gene sequences of the pyrgomorphid grasshopper genes, S. gregaria and D. melanogaster. The alignment shown assumes that two independent amino acid insertion occured in the gut copy of the pyrgomorphid species (yellow gaps in the other species). This hypothesis on the position of insertions maintains the conserved asparagine (N) at position 122 in the P. aegrotus gut copy and leads to the insertion of a histidine (H) after position 122. Numbering indicates the homologous positions in the pig Na,K-ATPase, transmembrane domains are colored in light green and numbered M1 - M6 (annotation of the crystal structure of the pig Na,K-ATPase 4HYT Protein Data Bank, [1]). Vertical red lines on top of the amino acids indicate that mutations at these sites have been functionally tested to increase resistance to cardiac glycosides in the vertebrate or insect enzyme ([2-15]), residues in red highlight exchanges at these positions and the two assumed amino acids insertions. NT - nervous tissue.