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cne23319-sup-0001-suppfig1.tif8876KSupplementary Figure 1 (This is a magenta-green version of Figure 2 from the manuscript for the assistance of color-blind readers.): The mature pattern of innervation on a partial segment of a leech heart tube. A: The mature presynaptic axons labeled by anti-ACT (green), is characterized by complex branching and extensive peptide-positive varicosities labeled by an anti-FMRFamide (red) label (presented here as magenta). FMRFamide appears to be preferentially located in varicosities devoid of microtubules. The arrow points to a main nerve that is projected by an HE neuron. Since anti-ACT labels all axons, many non-HE processes can be seen coursing through the body at different focal planes. The outline of the heart tube and side vessels (LaV and LdV) is indicated by dotted lines. B: Channels of labeled axons and varicosities that were independently traced and merged. The resultant pixel numbers were used to calculate an arbor index (R/ R+G+Y), which allowed us to estimate the total proportion of arbor length that is FMRF+ and ACT- in defined regions. Anterior is up and scale bar applies to both panels. (A magenta-green version of this figure has been posted as a supplementary file for the assistance of color-blind readers.)

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