Supplementary Fig. S10.

Ultrastructural localisation of secreted from muscle stage larvae (SML)-3. This figure shows a colour version of Fig. 7. (A) Formalin-fixed paraffin embedded Trichinella spiralis-infected rat tissue 28 days p.i. probed with anti-SML-3 and detected with horseradish peroxidase-conjugated anti-IgG and 3,3′-diaminobenzidine substrate. Sections were counterstained with Mayer’s haematoxylin. The black arrows show areas of staining within the mL1s stichocytes. Sections of methanol-fixed frozen T. spiralis infected rat tissue (24 days p.i.) were probed with anti-SML-3 and protein visualised with tetramethyl rhodamine iso-thiocyanate (TRITC)-conjugated anti-IgG. Nuclei within the tissue were visualised with DAPI. (B) Overlay of a TRITC (red) and DAPI (blue) images and localisation of SML-3 within the nematode (green arrows) but not the nurse cell. A 20 μm black (A) or 40 μm white (B) scale bar is shown in each image. Transmission electron microscopy sections of T. spiralis mL1s were probed with anti-SML-3 (C) and visualised with goat anti-mouse IgG coupled to 15 nm gold particles (black dots). Localisation of SML-3 to granules within the mL1s β-stichocytes (white arrows) can be observed. No staining of α-stichocyte granules was observed. Scale bar = 2 μm. αSt: α-stichocyte granules and βSt: β-stichocyte granules. Additional images, single channel images and pre-bleed controls are shown in Supplementary Figs. S11–S14.