Figure 5.

Distribution of myosin II, LMM, and Gal-GalNAc lectin in E. histolytica transfected with LMM. Transfectants growing in 30 μg/ml G418 amoebae were fixed and stained for myosin II, VSV-LMM, and Gal-GalNAc lectin. Confocal laser scanning microscopy was performed to detect immunofluorescence. Micrograph A shows rounded cells with myosin II labeling the plasma membrane (panel 1); some discrete myosin II spots are also observed in the cytoplasm. In the same cell, LMM is essentially diffuse throughout the cytoplasm (panel 2). The focal plane shown in this micrograph represents the middle of the cell. Micrograph B shows myosin II distribution in rounded cells (panel 3) and Gal-GalNAc in the same cell (panel 4). Myosin II is located under the plasma membrane as described previously. The Gal-GalNAc lectin is present at the membrane level; some patches of receptor can be seen (arrows). Micrograph C shows transfected amoeba that display a uroid (arrowhead). Myosin II is recruited to and concentrates in the uroid (panel 5); VSV-LMM is also observed in the uroid (panel 6). These results indicate the recruitment of both proteins during capping. Bar, 5 μm.