Figure 4

Immunolocalisation of FRePs in D. marginatus organs using fluorescence microscopy. Thin sections of midgut (C, F, H, I), salivary glands (A, B, J, K, L), and haemocytes attached to salivary glands (D, E, G) from fed female D. marginatus were labelled with sera raised against D. marginatus FRePs. Anti-DMF1 (A, B, C, D, E), anti-DMF2 (F, G), and anti-DMF3 (H, I, J, K) antibodies were used. Control reaction were carried out, in which anti-DMF sera were omitted (L). FRePs were localised on the surface and inside epithelial cells surrounding secretory granular cells of the acini type II (A, D, E); in granules of secretory cells located in the acini type III (B) and in haemocytes attached to the surface of salivary glands (D, E, G; arrows). Positive labelling reactions were observed in the cytoplasm of epithelium of salivary duct cells (J, K; arrows) and in thin cuticular layer facing to the lumen of the salivary duct (J, K). Anti-DMF sera detected structures on the surface (H) and inside the midgut cells (C, F, I). A, B, C, F, H, I, J, K, and L - Cy3-conjugated secondary antibodies (red); D, E, and G - FITC conjugated secondary antibodies (green). Cell nuclei were counter-stained using DAPI (blue).