Fig. 1

Migration of gp82-expressing T. cruzi metacyclic forms through the gastric mucin layer. a Transwell filters coated with gastric mucin were placed onto wells containing the indicated parasite strains. Samples were collected from the filter chamber at 30 min and 60 min for parasite counting. Values are the means ± SD of three independent experiments. As compared to CL strain, a significant lower migration of TcI strains (**P < 0.0005) and TcIV strain 1434 (*P < 0.005) was observed at 60 min. b Metacyclic forms, untreated (−) or treated (+) with 2 mg/ml pepsin, at pH 3.5, were analyzed by Western blot using mAb 3 F6 directed to gp82. As control of pepsin activity, BSA stained by Coomassie blue is shown. c Live parasites were incubated on ice for 1 h, in absence or in the presence of mAb 3 F6. After fixation, the parasites were incubated with Alexa Fluor 488-conjugated anti-IgG and the number of fluorescent parasites was estimated. d Parasites were incubated for 1 h in PBS. After centrifugation to remove parasites, the conditioned PBS was analyzed by Western blotting using mAb 3 F6 and mAb 5E7, directed to gp82 and gp90 respectively. e Transwell filters coated with gastric mucin were placed onto wells containing metacyclic forms of strain 515 alone, or in the presence of anti-gp90 mAb 5E7 that does not recognize live parasites or unrelated mAb 2C2. After 30 and 60 min incubation, samples from the filter chamber were taken for parasite counting. Values are the means ± variation of duplicates