Fig. 3.

Identification of in vitro secreted SmVAL4 and visualization of PNA-stained acetabular glands of parasites cultured for up to 7 days. Polyclonal anti-rSmVAL4 was used to identify native SmVAL4 in total parasite extracts (PE) and secretions containing released proteins (RP) of cercariae (Cerc), 3-hour schistosomula (3-h), day 3 schistosomula (3-day), day 5 schistosomula (5-day), and day 7 schistosomula (7-day). P1 and P2 - positive controls, recombinant SmVAL4, 15 and 50 ng, respectively. For PE, 10 μg of protein extract was applied in each lane a. Secretions derived from ~80.000 cultured schistosomula (20 μl) were used in b. Molecular weight (kDa) is marked on the left side of the images. c Parasite stages stained with Alexa-fluor 647-conjugated PNA (red). Green indicates FITC-phalloidin staining muscle fibers containing actin. Scale-bars: 50 μm