Figure 3.
The clathrin-adaptor protein GlENTHp participates in bLF and bLFcin uptake. (A) FITC-bLF and FITC-bLFcin are endocytosed to the PVs in wild-type and glenth–ha but not in ds-glenth trophozoites. The most representative effect is shown for each type of cell. Differential interference contrast (DIC) microscopy images are shown as insets. Bars, 5 μm. Representative image of wild-type and transgenic trophozoites showing the binding of FITC-bLF and FITC-bLFcin on the cell surface (including the flagella) when treated 10 min at 4 °C, washed and fixed with 4% formaldehyde (top insets). Histograms of fluorescent images (fluorescence/area × 103) show the relative amount of fluorescent bLF or bLFcin internalized by parasites in control (wild-type) and transgenic cells (glenth–ha and ds-glenth). All images were equally processed; the threshold value was determined and is exclusive in each image. The results are presented here as the average (±SD) of 10 determinations. (B) ds-glenth transgenic trophozoites treated with 12.5 μM of bLF or 2.6 μM of bLFcin show a non-deleterious effect on cell growth compared with glenth-ha cells. Data shown are representative of three independent experiments and are expressed as means of triplicates ***p-value < 0.001.