Expression pattern of putative carotenoid-binding protein genes. (A) Heat map of 24 takeout/juvenile hormone-binding protein (JHBP) family genes in P. xuthus. Red indicates positive values and green indicates negative values (color spectrum bar is shown to the right; N.D., not detected). Red circles indicate the genes examined by whole-mount in situ hybridization in (C). Stage and marking of each sample is shown above. The stage-specific co-expression cluster and marking specificity of each gene are also shown. See also Figures 2 to 4. (B) Neighbor joining tree of takeout/JHBP family genes based on their amino acid sequences. The numbers at the tree nodes represent the bootstrap values. The scale bars indicate the evolutionary distance between the groups. P. xuthus, D. plexippus and B. mori genes are shaded in red, blue and gray, respectively. Red asterisk indicates carotenoid-binding protein of locust Schistocerca gregaria, and black asterisks indicate takeout protein of Drosophila melanogaster and JHBP of Manduca sexta. Accession numbers are shown in parenthesis. Predicted genes of D. plexippus and B. mori were from MonarchBase http://monarchbase.umassmed.edu/ and KAIKObase http://sgp.dna.affrc.go.jp/KAIKObase/, respectively. (C) Spatial expression patterns of two putative carotenoid-binding protein mRNAs in P. xuthus and P. polytes larvae during the fourth and the third molts. Numbers in each panel indicate molt stage-hours after head capsule slippage. JHBP: juvenile hormone-binding protein.