Files in this Data Supplement:

• Supplemental Figure 1 (Adobe PDF) -

  Fig S1 Alignment of LWRh opsin sequences. The GenBank accession number for the full-length Vanessa cardui LWRh opsin also cloned in this study is AY613986.

• Supplemental Figure 2 (Adobe PDF) -

  Fig S2 Alignment of BRh1 and BRh2 opsin sequences. The glutamate at amino acid 107 confers blue-absorbing spectra (Salcedo et al., 2003).

• Supplemental Figure 3 (Adobe PDF) -

  Fig S3 Alignment of UVRh opsin sequences. The lysine at amino acid 112 confers UV-absorbing spectra (Salcedo et al., 2003). GenBank accession number for the full-length Vanessa cardui UVRh opsin also cloned in this study is AY613988.

• Supplemental Figure 4 (Adobe PDF) -

  Fig S4 Phylogeny of insect opsins. The tree was reconstructed using 299 amino acid sites and the neighbor-joining method with Poisson correction using 1000 bootstrap replicates. Numbers indicate percent bootstrap values. Only bootstrap support values >50% are shown. GenBank accession numbers are as follows: Anopheles gambiae (GPROP8, XM_312478; GPROP9, XM_319247); Antheraea pernyi (AB073299); Apis mellifera (AMUVOPS, AF004169; AMUVOPS, U70841; LWRh, U26026); Bicyclus anynana (UV, AF484248); Camponotus abdominalis (LW, U32502; SW, AF042788); Cataglyphis bombycina (LW, U32501; SW, AF042787); Danaus plexippus (Blue, AY605544; LW, AY605545; UV, AY605546); Drosophila melanogaster (Rh1, K02315; Rh2, M12896; Rh3, M17718; Rh4, NM_057353; Rh5, U67905; Rh6, Z86118); Lycaena rubidus (LWRh, AY587901; BRh1, AY587902; BRh2, AY587903; UVRh, AY587904); Manduca sexta (Manop1, L78080; Manop2, L78081; Manop3, AD001674); Megoura viciae (LW, AF189714; UV, AF189715); Papilio glaucus (PglRh1, AF077189; PglRh2, AF077190; PglRh3, AF067080; PglRh5, AF077191; PglRh6, AF077192); Papilio xuthus (PxRh1, AB007423; PxRh3, AB007425; PxRh4, AB028217; PxRh5, AB028218); Pieris rapae (PrB, AB208675; PrL, AB177984; PrUV, AB208673; PrV, AB208674); Schistocerca gregaria (Lo1, X80071; Lo2, X80072); Sphodromantis ssp. (X71665). Boxes indicate placement of L. rubidus UVRh (pink), BRh1 (dark blue), BRh2 (light blue) and LWRh (orange) sequences.

• Supplemental Tables -

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• Supplemental Figure 5 -

  Fig S5 Coordinate expression of BRh2 transcript with pink filtering pigment in dorsal eye. (A) Longitudinal section of the male retina showing coordinated expression of BRh2 with pink filtering pigment (arrow) (B) Tangential section of ommatidia showing the same coordinate expression of the pink filtering pigments with BRh2 mRNA.

• Supplemental Figure 6 -

  Fig S6 Electron micrographs of L. rubidus ommatidia. (A) Transverse section of a dorsal rim area ommatidium showing square-formed rhabdoms. The retinular cells are oriented vertically. (B) Transverse section of an ommatidium from the dorsal part of the retina in the median region of the retinula cell. A circular fused rhabdom is present. Note eight ambient tracheoles (tr). (C) Transverse section from the ventral domain of the retina in the same region of the retinula as in (B). Clusters of filtering pigment granules are seen in the R5-8 cells (arrowheads). (D) Transverse section of the biloped R9 receptor cell (asterisks). The cytoplasm is stained darker than in the other retinular cells (R1-R8). Arrow points to some microvilli of the rhabdom contributed by the basal R9 cell. (E) Longitudinal section of an ommatidium from the basal region of the main retina. The rhabdom shows a typical banded and interdigitating pattern, characteristic of ommatidia specialized for color vision.