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. 2015 Jul 31;11(7):e1005079. doi: 10.1371/journal.ppat.1005079

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© 2015 Avalos-Padilla et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 7 — (A) Western blot assays of trophozoites lysates from wild type clone A (lane A) and pNeo (lane pNeo) and pNeoEhvps32-HA (lane EhVps32) transfected cells, using the αrEhVps32 antibodies. As a loading control, the same membrane was re-blotted with αactin antibodies. (B) Densitometry analysis of bands showed in (A), normalized against actin band. (C) Diaminobenzidine-stained trophozoites that ingested erythrocytes for 30 min. (D) Rate of erythrophagocytosis of transfected trophozoites. (***) p<0.001. (E) Confocal microscopy of pNeoEhvps32-HA transfected trophozoites in resting conditions, using αrEhVps32 and αEhADH antibodies, followed by FITC-labeled and TRITC-labeled secondary antibodies. Arrows: membrane projections. (F-H) Trophozoites overexpressing EhVps32 incubated with erythrocytes for 2 min treated with αrEhVps32 and αEhADH antibodies and FITC- and TRITC-secondary antibodies, respectively. (F) Maximum projection of a transfected trophozoite evidencing the concentric arrays (arrow) formed by EhVps32. (G) Phase contrast. Arrow: concentric arrays. (H) Merging image (αrEhVps32 and αEhADH antibodies). Arrow: tunnel-like structure. e: erythrocytes.