Figure 3

Detection of FBA in Paracoccidioides . (A) Western blotting analysis. Different protein samples (15 μg) of Paracoccidioides comprehending the soluble and secreted proteins, cell wall fractions 1, 2 and 3 were obtained by sequential treatments as described in Materials and Methods. For negative and positive controls, we employed 3 μg of samples of bovine serum albumin (BSA) and the rFBA, respectively. The immunoblot was probed with the polyclonal antibodies directed to the rFBA. (B) Immunoelectron microscopy. Panel 1 - Transmission electron microscopy of Paracoccidioides yeast cells showing the cell wall (w), intracytoplasmic vacuoles (v), nucleus (n) and mitochondria (m). Panel 2 - Gold particles are observed in the cytoplasm region (arrows) and vesicles in release process (arrowheads). * corresponds to the region which has been expanded from panel 2. Panels 3 and 4 - Negative controls with anti-rabbit-igG-Au-conjugated and rabbit non immune sera, respectively. The bars indicate: 1.0 μm (Panel 1), 1.0 μm (Panel 2), 0.5 μm (Panel 3), 1.0 μm (Panel 4) and 0.5 μm (Zoom panel).