Labeling of Fal2-S1 transfectants with BODIPY-TR ceramide at trophozoite stage. In each panel, the first image (left to right) represents the bright field image, the second is the fluorescence signal from the GFP chimeric protein, third is an overlay of these two images, fourth is the BODIPY-TR fluorescence and fifth image is overlap of fluorescence by GFP and BODIPY-TR and sixth image is overlap of fluorescence by GFP, BODIPY-TR and DAPI. GFP fluorescence is seen overlapping with fluorescence by BODIPY-TR in the PV and food vacuole membrane. Some of the small foci having GFP fluorescence near PV (marked with solid arrows), show overlap with fluorescence by BODIPY-TR. Some of these foci in panel IV are seen fused with the food vacuole (marked with arrow head). In Fal2-S1 parasites, GFP fluorescence colocalized with BODIPY-TR-ceramide label, confirming the presence of fusion protein in food vacuole membrane. In many visual fields, the cytostomal vesicles containing GFP fusion proteins were found to be surrounded by BODIPY-Trceramide labeling suggesting that these foci are membrane bound structures (panels I and III).

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