Fig 7.

Immunolocalization of EhMSP-1 in E. histolytica trophozoites by confocal microscopy. (A) Western blot showing a single band in E. histolytica lysates detected using affinity-purified, rabbit polyclonal anti-EhMSP-1 IgG and an anti-rabbit IgG–HRP conjugate. (B) Immunofluorescent confocal microscopy showing control staining performed on detergent-permeabilized trophozoites using preimmune rabbit IgG. Trophozoites were revealed using an anti-SREHP monoclonal mouse IgG and an anti-mouse IgG–Alexa 488 conjugate (pseudocolored red). (C) Immunofluorescent confocal microscopy of permeabilized cells demonstrating the presence of EhMSP-1 in vesicles of all trophozoites. (D) Immunofluorescent confocal microscopy of nonpermeabilized trophozoites demonstrating the presence of EhMSP-1 on the surface of only a subset of amebas. EhMSP-1 was stained using affinity-purified anti-EhMSP-1 rabbit polyclonal IgG and an anti-rabbit IgG–Alexa 568 conjugate, and pseudocolored green. The SREHP was stained as for panel B. All scale bars shown are 10 μm.