Fig. 2

Immunostainings of Distal-less and Spalt proteins in larval and pupal wings. A–Dʹ Dll protein is present in late fifth instar larval and 24–26 h pupal wing discs. A, Aʹ, C and Cʹ In B. anynana larval and pupal wings, Dll is observed between veins as finger-like projections from the wing margin, ending with a discrete focus at the proximal tip of the fingers, that corresponds to the eyespot centres. In pupal stages of development, Dll becomes additionally observed in cells that correspond to the black scales of the eyespot pattern. B, Bʹ, D and Dʹ In P. canidia, intervein finger-like projections of Dll protein are observed but with no discrete foci at the tips of the fingers. E–Hʹ. Sal protein is present in late fifth instar larval and 24–26 h pupal wings discs. E, Eʹ, G and Gʹ In B. anynana, Sal protein is observed in eyespot foci during the larval stage. Like Dll, Sal becomes additionally observed in the cells that map to the black scales in the eyespots during pupal wing development. F, Fʹ, H and Hʹ In P. canidia, there is no cluster of cells in the middle of the spot pattern that is expressing higher levels of Sal proteins in larval wings, and Sal is present in the cells that map to the black scales in spots in 24 h pupal wings. I, Iʹ, J, Jʹ Dll and Sal proteins are also observed in cells that will become black scales located along the wing margin at both the wing tips and in the chevron patterns along the wing margin in P. canidia. Note the strong punctate nuclear staining of scale-building cells taken at 20× magnification. Scale bars for (C, D, G, and H—500 µm); (A, B, Bʹ, Cʹ, Dʹ, E, F, Gʹ and Hʹ—200 µm); (Eʹ and Fʹ—100 µm); (Aʹ, I, Iʹ, J and Jʹ)—50 µm)