Fig. 3. ACG protein in intact structures and cells. Intact wild-type slugs (A), fruiting bodies (B) and spores (C) were fixed in methanol and stained with ACG antibody and FITC-conjugated goat-anti-rabbit IgG (GARFITC). (D-F). Slugs were dissociated into single cells, which were first stained with ACG antibody and GARFITC, and subsequently with mouse monoclonal antibody mAb83.5 that was raised against the spore coat protein SP85 (Zhang et al., 1999) and Texas Red-conjugated goat-anti-mouse IgG. Intact structures and cells were photographed using a Leica TCS SP2 confocal laser scanning microscope, using 596 nm excitation and 620 nm emission for Texas Red, and 495 nm excitation and 520 nm emission for FITC. D and E are superimposed in F to show colocalization of ACG and SP85. Scale bars: A,B, 100 µm; C, 1 µm; D, 10 µm.